Microtubule organization within mitotic spindles revealed by serial block face scanning electron microscopy and image analysis

Serial block face scanning electron microscopy (SBF-SEM) is a powerful method to analyze cells in 3D. Here, working at the resolution limit of the method, we describe a correlative light-SBF-SEM workflow to resolve microtubules of the mitotic spindle in human cells. We present four examples of uses for this workflow that are not practical by light microscopy and/or transmission electron microscopy. First, distinguishing closely associated microtubules within K-fibers; second, resolving bridging fibers in the mitotic spindle; third, visualizing membranes in mitotic cells, relative to the spindle apparatus; and fourth, volumetric analysis of kinetochores. Our workflow also includes new computational tools for exploring the spatial arrangement of microtubules within the mitotic spindle. We use these tools to show that microtubule order in mitotic spindles is sensitive to the level of TACC3 on the spindle

Ice-sheet dynamics through the Quaternary on the mid-Norwegian continental margin inferred from 3D seismic data

Reconstructing the evolution of ice sheets is critical to our understanding of the global environmental system, but most detailed palaeo-glaciological reconstructions have hitherto focused on the very recent history of ice sheets. Here, we present a three-dimensional (3D) reconstruction of the changing nature of ice-sheet derived sedimentary architecture through the Quaternary Ice Age of almost 3 Ma. An extensive geophysical record documents a marine-terminating, calving Fennoscandian Ice Sheet (FIS) margin present periodically on the mid-Norwegian shelf since the beginning of the Quaternary. Spatial and temporal variability of the FIS is illustrated by the gradual development of fast-flowing ice streams and associated intensification of focused glacial erosion and sedimentation since that time. Buried subglacial landforms reveal a complex and dynamic ice sheet, with converging palaeo-ice streams and several flow-switching events that may reflect major changes in topography and basal thermal regime. Lack of major subglacial meltwater channels suggests a largely distributed drainage system beneath the marine-terminating part of the FIS. This palaeo-environmental examination of the FIS provides a useful framework for ice-sheet modelling and shows that fragmentary preservation of buried surfaces and variability of ice-sheet dynamics should be taken into account when reconstructing glacial history from spatially limited datasets.Doctoral funding for A.M. was from the Gates Cambridge Trust. Datasets were provided by the Norwegian Petroleum Directorate (NPD), and Norwegian University of Science & Technology (NTNU) from the Schlumberger Petrel Ready Database. Authors would like to thank Schlumberger® for the dataset and access to the Petrel Schlumberger® Software that was provided by NTNU Department of Geophysics and Petroleum Engineering

3D seismic evidence of buried iceberg ploughmarks from the mid-Norwegian continental margin reveals largely persistent North Atlantic Current through the Quaternary.

Over 7500 buried linear and curvilinear depressions interpreted as iceberg ploughmarks were identified within the Quaternary Naust Formation from an extensive three-dimensional seismic dataset that covers ~ 40,000 km2 of the mid-Norwegian continental margin. The morphology and net orientation of ploughmarks were mapped and analysed. These features are up to 28 km long, 700 m wide and are incised up to 31 m deep. On average, ploughmarks are incised 5 m deep, with median width of 185 m and median lengths ranging from 1.2 to 2.7 km for individual palaeo-surfaces. Width to depth ratio ranges from 8:1 to 400:1 and is on average 36:1. The presence of ploughmarks buried deeply within some palaeo-slope surfaces implies the occasional presence of very large icebergs since the middle Quaternary, suggesting that thick ice-sheet margins with fast-flowing ice streams were present in order to calve icebergs of such dimensions into the Norwegian Sea. The wide geographical distribution of ploughmarks suggests unrestricted iceberg drift and an open Norwegian Sea during the periods of iceberg calving since the early Quaternary. Ploughmark trajectory analysis demonstrates that the ocean current circulation, now dominated by the northeasterly flowing Norwegian Atlantic Current (NwAC), has largely persisted throughout the Quaternary. Despite the overall strikingly consistent pattern of iceberg drift, ploughmark mapping also shows evidence for short-lived NwAC reductions possibly related to major phases of iceberg discharge and/or meltwater pulses from the Fennoscandian Ice Sheet during the middle and late Quaternary

Efficient Parameter-Estimating Algorithms for Symmetry-Motivated Models: Econometrics and Beyond

It is known that symmetry ideas can explain the empirical success of many non-linear models. This explanation makes these models theoretically justified and thus, more reliable. However, the models remain non-linear and thus, identification or the model\u27s parameters based on the observations remains a computationally expensive nonlinear optimization problem. In this paper, we show that symmetry ideas can not only help to select and justify a nonlinear model, they can also help us design computationally efficient almost-linear algorithms for identifying the model\u27s parameters

Insights from extreme coral reefs in a changing world

© 2020, Springer-Verlag GmbH Germany, part of Springer Nature. Coral reefs are one of the most biodiverse and economically important ecosystems in the world, but they are rapidly degrading due to the effects of global climate change and local anthropogenic stressors. Reef scientists are increasingly studying coral reefs that occur in marginal and extreme environments to understand how organisms respond to, and cope with, environmental stress, and to gain insight into how reef organisms may acclimate or adapt to future environmental change. To date, there have been more than 860 publications describing the biology and/or abiotic conditions of marginal and extreme reef environments, most of which were published within the past decade. These include systems characterized by unusually high, low, and/or variable temperatures (intertidal, lagoonal, high-latitude areas, and shallow seas), turbid or urban environments, acidified habitats, and mesophotic depth, and focus on reefs geographically spread throughout most of the tropics. The papers in this special issue of Coral Reefs, entitled Coral Reefs in a Changing World: Insights from Extremes, build on the growing body of literature on these unique and important ecosystems, providing a deeper understanding of the patterns and processes governing life in marginal reef systems, and the implications that these insights may have for the future of tropical coral reefs in our rapidly changing world

Metabolic design of macroscopic bioreaction models: application to Chinese hamster ovary cells

The aim of this paper is to present a systematic methodology to design macroscopic bioreaction models for cell cultures based upon metabolic networks. The cell culture is seen as a succession of phases. During each phase, a metabolic network represents the set of reactions occurring in the cell. Then, through the use of the elementary flux modes, these metabolic networks are used to derive macroscopic bioreactions linking the extracellular substrates and products. On this basis, as many separate models are obtained as there are phases. Then, a complete model is obtained by smoothly switching from model to model. This is illustrated with batch cultures of Chinese hamster ovary cells

Pseudomonas aeruginosa Adaptation to Lungs of Cystic Fibrosis Patients Leads to Lowered Resistance to Phage and Protist Enemies

Pathogenic life styles can lead to highly specialized interactions with host species, potentially resulting in fitness trade-offs in other ecological contexts. Here we studied how adaptation of the environmentally transmitted bacterial pathogen, Pseudomonas aeruginosa, to cystic fibrosis (CF) patients affects its survival in the presence of natural phage (14/1, ΦKZ, PNM and PT7) and protist (Tetrahymena thermophila and Acanthamoebae polyphaga) enemies. We found that most of the bacteria isolated from relatively recently intermittently colonised patients (1-25 months), were innately phage-resistant and highly toxic for protists. In contrast, bacteria isolated from long time chronically infected patients (2-23 years), were less efficient in both resisting phages and killing protists. Moreover, chronic isolates showed reduced killing of wax moth larvae (Galleria mellonella) probably due to weaker in vitro growth and protease expression. These results suggest that P. aeruginosa long-term adaptation to CF-lungs could trade off with its survival in aquatic environmental reservoirs in the presence of microbial enemies, while lowered virulence could reduce pathogen opportunities to infect insect vectors; factors that are both likely to result in poorer environmental transmission. From an applied perspective, phage therapy could be useful against chronic P. aeruginosa lung infections that are often characterized by multidrug resistance: chronic isolates were least resistant to phages and their poor growth will likely slow down the emergence of beneficial resistance mutations

Change in Composition of the Anopheles Gambiae Complex and its Possible Implications for the Transmission of Malaria and Lymphatic Filariasis in North-Eastern Tanzania.

A dramatic decline in the incidence of malaria due to Plasmodium falciparum infection in coastal East Africa has recently been reported to be paralleled (or even preceded) by an equally dramatic decline in malaria vector density, despite absence of organized vector control. As part of investigations into possible causes for the change in vector population density, the present study analysed the Anopheles gambiae s.l. sibling species composition in north-eastern Tanzania. The study was in two parts. The first compared current species complex composition in freshly caught An. gambiae s.l. complex from three villages to the composition reported from previous studies carried out 2-4 decades ago in the same villages. The second took advantage of a sample of archived dried An. gambiae s.l. complex specimens collected regularly from a fourth study village since 2005. Both fresh and archived dried specimens were identified to sibling species of the An. gambiae s.l. complex by PCR. The same specimens were moreover examined for Plasmodium falciparum and Wuchereria bancrofti infection by PCR. As in earlier studies, An. gambiae s.s., Anopheles merus and Anopheles arabiensis were identified as sibling species found in the area. However, both study parts indicated a marked change in sibling species composition over time. From being by far the most abundant in the past An. gambiae s.s. was now the most rare, whereas An. arabiensis had changed from being the most rare to the most common. P. falciparum infection was rarely detected in the examined specimens (and only in An. arabiensis) whereas W. bancrofti infection was prevalent and detected in all three sibling species. The study indicates that a major shift in An. gambiae s.l. sibling species composition has taken place in the study area in recent years. Combined with the earlier reported decline in overall malaria vector density, the study suggests that this decline has been most marked for An. gambiae s.s., and least for An. arabiensis, leading to current predominance of the latter. Due to differences in biology and vectorial capacity of the An. gambiae s.l. complex the change in sibling species composition will have important implications for the epidemiology and control of malaria and lymphatic filariasis in the study area