Human Naive Pluripotent Stem Cells Model X Chromosome Dampening and X Inactivation

Naive human embryonic stem cells (hESCs) can be derived from primed hESCs or directly from blastocysts, but their X chromosome state has remained unresolved. Here, we show that the inactive X chromosome (Xi) of primed hESCs was reactivated in naive culture conditions. Like cells of the blastocyst, the resulting naive cells contained two active X chromosomes with XIST expression and chromosome-wide transcriptional dampening and initiated XIST-mediated X inactivation upon differentiation. Both establishment of and exit from the naive state (differentiation) happened via an XIST-negative XaXaintermediate. Together, these findings identify a cell culture system for functionally exploring the two X chromosome dosage compensation processes in early human development: X dampening and X inactivation. However, remaining differences between naive hESCs and embryonic cells related to mono-allelic XIST expression and non-random X inactivation highlight the need for further culture improvement. As the naive state resets Xiabnormalities seen in primed hESCs, it may provide cells better suited for downstream applications

Comparison of reprogramming factor targets reveals both species-specific and conserved mechanisms in early iPSC reprogramming

Abstract Background Both human and mouse fibroblasts can be reprogrammed to pluripotency with Oct4, Sox2, Klf4, and c-Myc (OSKM) transcription factors. While both systems generate pluripotency, human reprogramming takes considerably longer than mouse. Results To assess additional similarities and differences, we sought to compare the binding of the reprogramming factors between the two systems. In human fibroblasts, the OSK factors initially target many more closed chromatin sites compared to mouse. Despite this difference, the intra- and intergenic distribution of target sites, target genes, primary binding motifs, and combinatorial binding patterns between the reprogramming factors are largely shared. However, while many OSKM binding events in early mouse cell reprogramming occur in syntenic regions, only a limited number is conserved in human. Conclusions Our findings suggest similar general effects of OSKM binding across these two species, even though the detailed regulatory networks have diverged significantly

Proteomic and protein interaction network analysis of human T lymphocytes during cell-cycle entry

Proteomic analysis of T cells emerging from quiescence identifies dynamic network-level changes in key cellular processes. Disruption of two such processes, ribosome biogenesis and RNA splicing, reveals that the programs controlling cell growth and cell-cycle entry are separable

Lightning Jump Algorithm Development for the GOESR Geostationary Lightning Mapper

Current work on the lightning jump algorithm to be used in GOESR Geostationary Lightning Mapper (GLM)'s data stream is multifaceted due to the intricate interplay between the storm tracking, GLM proxy data, and the performance of the lightning jump itself. This work outlines the progress of the last year, where analysis and performance of the lightning jump algorithm with automated storm tracking and GLM proxy data were assessed using over 700 storms from North Alabama. The cases analyzed coincide with previous semiobjective work performed using total lightning mapping array (LMA) measurements in Schultz et al. (2011). Analysis shows that key components of the algorithm (flash rate and sigma thresholds) have the greatest influence on the performance of the algorithm when validating using severe storm reports. Automated objective analysis using the GLM proxy data has shown probability of detection (POD) values around 60% with false alarm rates (FAR) around 73% using similar methodology to Schultz et al. (2011). However, when applying verification methods similar to those employed by the National Weather Service, POD values increase slightly (69%) and FAR values decrease (63%). The relationship between storm tracking and lightning jump has also been tested in a realtime framework at NSSL. This system includes fully automated tracking by radar alone, realtime LMA and radar observations and the lightning jump. Results indicate that the POD is strong at 65%. However, the FAR is significantly higher than in Schultz et al. (2011) (5080% depending on various tracking/lightning jump parameters) when using storm reports for verification. Given known issues with Storm Data, the performance of the realtime jump algorithm is also being tested with high density radar and surface observations from the NSSL Severe Hazards Analysis & Verification Experiment (SHAVE)

SPA: Economical and workload-driven indexing for data analytics in the cloud

Selective queries are not uncommon in large-scale data analytics, for example, when drilling down into a specific customer in a dashboard. Traditionally, selective queries are accelerated by creating secondary indexes. However, because of their large size, expensive maintenance, and difficulty to tune and automate, indexes are typically not used in modern cloud data warehouses or data lakes. Instead, such systems rely mostly on full table scans and lightweight optimizations like min/max filtering, whose effectiveness depends heavily on the data layout and value distributions.We propose SPA as the vision for automatically optimizing selective queries for immutable copy-on-write data formats. SPA adaptively indexes subsets of the data in an incremental and workload-driven manner. It makes fine-grained decisions and continuously monitors their benefit, dynamically allocating an optimization budget in a way that bounds the additional cost of indexing. Furthermore, it guarantees a performance improvement in the cases where indexes - potentially partial ones - prove to be beneficial. When indexes lose their benefit due to a shifting workload, they are gradually deconstructed in favor of optimizations that accommodate recent trends. As SPA does not require information about updates performed on the data, it can also be employed as an accelerator for systems that do not control the data, e.g., in cloud data lakes

An Image-Free Opto-Mechanical System for Creating Virtual Environments and Imaging Neuronal Activity in Freely Moving Caenorhabditis elegans

Non-invasive recording in untethered animals is arguably the ultimate step in the analysis of neuronal function, but such recordings remain elusive. To address this problem, we devised a system that tracks neuron-sized fluorescent targets in real time. The system can be used to create virtual environments by optogenetic activation of sensory neurons, or to image activity in identified neurons at high magnification. By recording activity in neurons of freely moving C. elegans, we tested the long-standing hypothesis that forward and reverse locomotion are generated by distinct neuronal circuits. Surprisingly, we found motor neurons that are active during both types of locomotion, suggesting a new model of locomotion control in C. elegans. These results emphasize the importance of recording neuronal activity in freely moving animals and significantly expand the potential of imaging techniques by providing a mean to stabilize fluorescent targets

Packages of Care for Attention-Deficit Hyperactivity Disorder in Low- and Middle-Income Countries

In the sixth in a series of six articles on packages of care for mental disorders in low- and middle-income countries, Alan Flisher and colleagues discuss the treatment of attention-deficit hyperactivity disorder

A microfluidics-based method for measuring neuronal activity in Drosophila chemosensory neurons

Monitoring neuronal responses to defined sensory stimuli is a powerful and widely used approach for understanding sensory coding in the nervous system. However, providing precise, stereotypic and reproducible cues while concomitantly recording neuronal activity remains technically challenging. Here we describe the fabrication and use of a microfluidics system that allows precise temporally restricted stimulation of Drosophila chemosensory neurons with an array of different chemical cues. The system can easily be combined with genetically encoded calcium sensors, and it can measure neuronal activity at single-cell resolution in larval sense organs and in the proboscis or leg of the adult fly. We describe the design of the master mold, the production of the microfluidic chip and live imaging using the calcium sensor GCaMP, expressed in distinct types of Drosophila chemosensory neurons. Fabrication of the master mold and microfluidic chips requires basic skills in photolithography and takes ~2 weeks; the same devices can be used repeatedly over several months. Flies can be prepared for measurements in minutes and imaged for up to 1 h