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Candida albicans displays a variety of morphological forms, and the ability to switch forms must be linked with cell cycle control. In budding yeast the Mitotic Exit Network (MEN) acts to drive mitotic exit and signal for cytokinesis and cell separation. However, previous reports on the MEN in C . albicans have raised questions on its role in this organism, with the components analysed to date demonstrating differing levels of importance in the processes of mitotic exit, cytokinesis and cell separation. This work focuses on the role of the Cdc15 kinase in C . albicans and demonstrates that, similar to Saccharomyces cerevisiae , it plays an essential role in signalling for mitotic exit and cytokinesis. Cells depleted of Cdc15 developed into elongated filaments, a common response to cell cycle arrest in C . albicans . These filaments emerged exclusively from large budded cells, contained two nuclear bodies and exhibited a hyper-extended spindle, all characteristic of these cells failing to exit mitosis. Furthermore these filaments displayed a clear cytokinesis defect, and CDC15 over-expression led to aberrant cell separation following hyphal morphogenesis. Together, these results are consistent with Cdc15 playing an essential role in signalling for mitotic exit, cytokinesis and cell separation in C . albicans .

Monitoring the structure of forest stands is of high importance for forest managers to help them in maintaining ecosystem services. For that purpose, Unmanned Aerial Vehicles (UAVs) open new prospects, especially in combination with Light Detection and Ranging (LiDAR) technology. Indeed, the shorter distance from the Earth’s surface significantly increases the point density beneath the canopy, thus offering new possibilities for the extraction of the underlying semantics. For example, tree stems can now be captured with sufficient detail, which is a gateway to accurately locating trees and directly retrieving metrics—e.g., the Diameter at Breast Height (DBH). Current practices usually require numerous site-specific parameters, which may preclude their use when applied beyond their initial application context. To overcome this shortcoming, the machine learning Hierarchical Density-Based Spatial Clustering of Application of Noise (HDBSCAN) clustering algorithm was further improved and implemented to segment tree stems. Afterwards, Principal Component Analysis (PCA) was applied to extract tree stem orientation for subsequent DBH estimation. This workflow was then validated using LiDAR point clouds collected in a temperate deciduous closed-canopy forest stand during the leaf-on and leaf-off seasons, along with multiple scanning angle ranges. The results show that the proposed methodology can correctly detect up to 82% of tree stems (with a precision of 98%) during the leaf-off season and have a Maximum Scanning Angle Range (MSAR) of 75 degrees, without having to set up any site-specific parameters for the segmentation procedure. In the future, our method could then minimize the omission and commission errors when initially detecting trees, along with assisting further tree metrics retrieval. Finally, this research shows that, under the study conditions, the point density within an approximately 1.3-meter height above the ground remains low within closed-canopy forest stands even during the leaf-off season, thus restricting the accurate estimation of the DBH. As a result, autonomous UAVs that can both fly above and under the canopy provide a clear opportunity to achieve this purpose.

The Mesozoic Era is characterized by numerous oceanic anoxic events (OAEs) that are diagnostically expressed by widespread marine organic-carbon burial and coeval carbon-isotope excursions. Here we present coupled high-resolution carbon- and sulfur-isotope data from four European OAE 2 sections spanning the Cenomanian–Turonian boundary that show roughly parallel positive excursions. Significantly, however, the interval of peak magnitude for carbon isotopes precedes that of sulfur isotopes with an estimated offset of a few hundred thousand years. Based on geochemical box modeling of organic-carbon and pyrite burial, the sulfur-isotope excursion can be generated by transiently increasing the marine burial rate of pyrite precipitated under euxinic (i.e., anoxic and sulfidic) water-column conditions. To replicate the observed isotopic offset, the model requires that enhanced levels of organic-carbon and pyrite burial continued a few hundred thousand years after peak organic-carbon burial, but that their isotope records responded differently due to dramatically different residence times for dissolved inorganic carbon and sulfate in seawater. The significant inference is that euxinia persisted post-OAE, but with its global extent dwindling over this time period. The model further suggests that only ∼5% of the global seafloor area was overlain by euxinic bottom waters during OAE 2. Although this figure is ∼30× greater than the small euxinic fraction present today (∼0.15%), the result challenges previous suggestions that one of the best-documented OAEs was defined by globally pervasive euxinic deep waters. Our results place important controls instead on local conditions and point to the difficulty in sustaining whole-ocean euxinia.

Shed antler hunting (i.e., collecting cast cervid antlers) has increased in popularity during the past decade, but little is known about how this recreational activity affects ungulate movements and space use. We placed geographic positioning system (GPS)-collars on 133 female and male bighorn sheep (Ovis canadensis), bison (Bison bison), and mule deer (Odocoileus hemionus) to quantify their movements and space use during shed antler hunts compared with those behaviors during helicopter surveys in Utah, USA, from 2012 to 2015. For each species, we calculated means and 95% confidence intervals for distance moved during 90-minute segments (16 points/day) pre-event (control, 7 consecutive days prior to event), event (1–2 days), and post-event (7 consecutive days after event) for shed antler hunts and helicopter surveys. We also compared use of space for each species during these events. Female bighorn sheep did not increase distance moved or substantially change space use during shed antler hunts and helicopter surveys. Male bighorn sheep increased distance moved 41% on average during shed antler hunts and by 2.02 times during helicopter surveys but did not change space use during those events. Female bison increased distance moved 15% on average during shed antler hunts and 30% during helicopter surveys. Mule deer increased distance moved and altered space use the most during shed antler hunts; females increased distance moved 97%, and 54% of females moved a mean distance of 742±642 (SD) m (range=9–3,778 m) outside of their home ranges during those hunts for a mean of 9.2±9.4 hours (range=1.5 to 41 hr). Male mule deer increased distance moved by 2.10 times on average during shed antler hunts, and 82% of males moved a mean distance of 1,264±732m (range=131–3,637 m) outside of their home ranges during those hunts for a mean of 12.6±7.6 hours (range=4.5–33 hr). Our results provide timely information about how legal shed antler hunting affects movements and space use of female and male ungulates, especially mule deer, and can guide the conservation of ungulate populations and their habitat.

Continental interiors were flooded by epeiric seas during many intervals of the geologic past. Few modern analogs exist for these environments, however, and basic variables such as redox, salinity, and restriction are difficult to reconstruct in deep time. Despite these challenges, constraining epeiric watermass properties is critical because much of our preserved and accessible sedimentary record was deposited in such settings. Here, we present a four‐dimensional reconstruction of watermass evolution in the Late Devonian Appalachian Seaway of North America. We use combined proxies for sediment supply, paleosalinity, paleoredox, and basin hydrography in six cores through the Upper Devonian Cleveland Shale deposited across a paleo‐depth transect. Cyclic, coupled changes in sedimentation, redox, and salinity are recorded in environments near the Catskill Delta. Additionally, a pronounced salinity gradient was present from low‐brackish conditions near the delta to fully marine conditions in the basin interior, with a lower‐salinity mixing zone recorded across the Cumberland Sill. We also identified two broad sequences—the lower and upper Cleveland Shale—each of which shows distinct watermass signatures. The lower Cleveland Shale records a redox gradient with euxinia only present along the Cumberland Sill, whereas the upper Cleveland Shale records intensification of euxinia (potentially in the photic zone) at all six sites, which may be coincident with the Hangenberg extinction event. Ultimately, this study identifies pronounced epeiric watermass gradients over short timescales (millennia) and distances (hundreds of km or less), highlighting the need for interpreting the geochemistry of epicontinental deposits in the context of basin hydrography and paleosalinity. Plain Language Summary The interiors of continents were flooded by shallow seas during many intervals of Earth history; however, little is known about the basic watermass properties of ancient inland seas and how they differed from the open ocean. Here, we provide an example of watermass reconstruction in an ancient inland sea by investigating the Cleveland Shale, which was deposited in a flooded region west of the Appalachian Mountains during the Late Devonian (∼383–359 million years ago). We use geochemical proxies for oxygen concentrations (redox) and salinity in six rock cores located across a gradient of ancient water depth. Our data reveal a pronounced salinity gradient and two discrete stages in the sea's chemical evolution, with the first stage recording a strong redox gradient across the sea and the second stage characterized by oxygen deficiency and toxic hydrogen sulfide at all sites investigated. This expansion of shallow‐water hydrogen sulfide occurred at the same time as the Hangenberg mass extinction event that killed many inhabitants of the shallow oceans. Ultimately, this study is among the first to identify pronounced chemical gradients in an ancient inland sea and link spatiotemporal watermass properties to ocean habitability and extinction during a critical time in Earth history. Key Points Four‐dimensional paleoredox, paleosalinity, and hydrographic reconstruction of the Late Devonian Appalachian Seaway Pronounced watermass heterogeneity over short timescales and distances, including large vertical and lateral salinity gradient Basin‐wide intensification of water column (and potentially photic zone) euxinia during the Hangenberg extinction event

To comply with guiding principles for the ethical use of animals for experimental research, the field of mutation research has witnessed a shift of interest from large-scale in vivo animal experiments to small-sized in vitro studies. Mutation assays in cultured cells of transgenic rodents constitute, in many ways, viable alternatives to in vivo mutagenicity experiments in the corresponding animals. A variety of transgenic rodent cell culture models and mutation detection systems have been developed for mutagenicity testing of carcinogens. Of these, transgenic Big Blue® (Stratagene Corp., La Jolla, CA, USA, acquired by Agilent Technologies Inc., Santa Clara, CA, USA, BioReliance/Sigma-Aldrich Corp., Darmstadt, Germany) mouse embryonic fibroblasts and the λ Select cII Mutation Detection System have been used by many research groups to investigate the mutagenic effects of a wide range of chemical and/or physical carcinogens. Here, we review techniques and principles involved in preparation and culturing of Big Blue® mouse embryonic fibroblasts, treatment in vitro with chemical/physical agent(s) of interest, determination of the cII mutant frequency by the λ Select cII assay and establishment of the mutation spectrum by DNA sequencing. We describe various approaches for data analysis and interpretation of the results. Furthermore, we highlight representative studies in which the Big Blue® mouse cell culture model and the λ Select cII assay have been used for mutagenicity testing of diverse carcinogens. We delineate the advantages of this approach and discuss its limitations, while underscoring auxiliary methods, where applicable.

Monitoring of canopy density with related metrics such as leaf area index (LAI) makes a significant contribution to understanding and predicting processes in the soil–plant–atmosphere system and to indicating crop health and potential yield for farm management. Remote sensing methods using optical sensors that rely on spectral reflectance to calculate LAI have become more mainstream due to easy entry and availability. Methods with vegetation indices (VI) based on multispectral reflectance data essentially measure the green area index (GAI) or response to chlorophyll content of the canopy surface and not the entire aboveground biomass that may be present from non-green elements that are key to fully assessing the carbon budget. Methods with light detection and ranging (LiDAR) have started to emerge using gap fraction (GF) to estimate the plant area index (PAI) based on canopy density. These LiDAR methods have the main advantage of being sensitive to both green and non-green plant elements. They have primarily been applied to forest cover with manned airborne LiDAR systems (ALS) and have yet to be used extensively with crops such as winter wheat using LiDAR on unmanned aircraft systems (UAS). This study contributes to a better understanding of the potential of LiDAR as a tool to estimate canopy structure in precision farming. The LiDAR method proved to have a high to moderate correlation in spatial variation to the multispectral method. The LiDAR-derived PAI values closely resemble the SunScan Ceptometer GAI ground measurements taken early in the growing season before major stages of senescence. Later in the growing season, when the canopy density was at its highest, a possible overestimation may have occurred. This was most likely due to the chosen flight parameters not providing the best depictions of canopy density with consideration of the LiDAR’s perspective, as the ground-based destructive measurements provided lower values of PAI. Additionally, a distinction between total LiDAR-derived PAI, multispectral-derived GAI, and brown area index (BAI) is made to show how the active and passive optical sensor methods used in this study can complement each other throughout the growing season.

The potential for human disease treatment using human pluripotent stem cells, including embryonic stem cells and induced pluripotent stem cells (iPSCs), also carries the risk of added genomic instability. Genomic instability is most often linked to DNA repair deficiencies, which indicates that screening/characterization of possible repair deficiencies in pluripotent human stem cells should be a necessary step prior to their clinical and research use. In this study, a comparison of DNA repair pathways in pluripotent cells, as compared to those in non-pluripotent cells, demonstrated that DNA repair capacities of pluripotent cell lines were more heterogeneous than those of differentiated lines examined and were generally greater. Although pluripotent cells had high DNA repair capacities for nucleotide excision repair, we show that ultraviolet radiation at low fluxes induced an apoptotic response in these cells, while differentiated cells lacked response to this stimulus, and note that pluripotent cells had a similar apoptotic response to alkylating agent damage. This sensitivity of pluripotent cells to damage is notable since viable pluripotent cells exhibit less ultraviolet light-induced DNA damage than do differentiated cells that receive the same flux. In addition, the importance of screening pluripotent cells for DNA repair defects was highlighted by an iPSC line that demonstrated a normal spectral karyotype, but showed both microsatellite instability and reduced DNA repair capacities in three out of four DNA repair pathways examined. Together, these results demonstrate a need to evaluate DNA repair capacities in pluripotent cell lines, in order to characterize their genomic stability, prior to their pre-clinical and clinical use.

In the past decade, Galleria mellonella (wax moth) larvae have become widely used as a non-mammalian infection model. However, the full potential of this infection model has yet to be realised, limited by the variable quality of larvae used and the lack of standardised procedures. Here, we review larvae suitable for research, protocols for dosing larvae, and methods for scoring illness in larvae infected with fungal pathogens. The development of standardised protocols for carrying out our experimental work will allow high throughput screens to be developed, changing the way in which we evaluate panels of mutants and strains. It will also enable the in vivo screening of potential antimicrobials at an earlier stage in the research and development cycle.

Environmental chemicals and radiation have often been implicated in producing alterations of the epigenome thus potentially contributing to cancer and other diseases. Ionizing radiation, released during accidents at nuclear power plants or after atomic bomb explosions, is a potentially serious health threat for the exposed human population. This type of high-energy radiation causes DNA damage including single- and double-strand breaks and induces chromosomal rearrangements and mutations, but it is not known if ionizing radiation directly induces changes in the epigenome of irradiated cells. We treated normal human fibroblasts and normal human bronchial epithelial cells with different doses of γ-radiation emitted from a cesium 137 ((137)Cs) radiation source. After a seven-day recovery period, we analyzed global DNA methylation patterns in the irradiated and control cells using the methylated-CpG island recovery assay (MIRA) in combination with high-resolution microarrays. Bioinformatics analysis revealed only a small number of potential methylation changes with low fold-difference ratios in the irradiated cells. These minor methylation differences seen on the microarrays could not be verified by COBRA (combined bisulfite restriction analysis) or bisulfite sequencing of selected target loci. Our study shows that acute γ-radiation treatment of two types of human cells had no appreciable direct effect on DNA cytosine methylation patterns in exposed cells.