Miracle-dz/UniFAR-FTUs huggingface dataset:
High throughput single-cell genome sequencing gives insights into the generation and evolution of mosaic aneuploidy in Leishmania donovani

Leishmania, a unicellular eukaryotic parasite, is a unique model for aneuploidy and cellular heterogeneity, along with their potential role in adaptation to environmental stresses. Somy variation within clonal populations was previously explored in a small subset of chromosomes using fluorescence hybridization methods. This phenomenon, termed mosaic aneuploidy (MA), might have important evolutionary and functional implications but remains under-explored due to technological limitations. Here, we applied and validated a high throughput single-cell genome sequencing method to study for the first time the extent and dynamics of whole karyotype heterogeneity in two Leishmania clonal populations representing different stages of MA evolution in vitro. We found that drastic changes in karyotypes quickly emerge in a population stemming from an almost euploid founder cell. This possibly involves polyploidization/hybridization at an early stage of population expansion, followed by assorted ploidy reduction. During further stages of expansion, MA increases by moderate and gradual karyotypic alterations. MA usually affected a defined subset of chromosomes, of which some display an enrichment in snoRNA genes which could represent an adaptative benefit to the amplification of these chromosomes. Our data provide the first complete characterization of MA in Leishmania and pave the way for further functional studies. Note to the BioRxiv community The present preprint is a revision of an older preprint posted on 06th March 2020 on BioRxiv (https://www.biorxiv.org/content/10.1101/2020.03.05.976233v1). Here we included two extra samples in our single-cell genome sequencing (SCGS) analysis – the BPK081 cl8 clone (a nearly euploid strain) and a population consisting of a mixture of four L. donovani strains which was used as control for high levels of mosaicism in aneuploidy and for estimation of doublets. We also upgraded the bioinformatics pipeline to determine single-cell karyotypes and performed new fluorescence in situ hybridization (FISH) analysis. The new findings observed especially in the BPK081 cl8 led to a reformulation of the text, a new hypothesis for the evolution of mosaicism and a general restructuring of the article. Therefore, the older preprint is obsolete.
Preclinical evaluation of a SARS-CoV-2 mRNA vaccine PTX-COVID19-B

Safe and effective vaccines are needed to end the COVID-19 pandemic caused by SARS-CoV-2. Here we report the preclinical development of a lipid nanoparticle (LNP) formulated SARS-CoV-2 mRNA vaccine, PTX-COVID19-B. PTX-COVID19-B was chosen among three candidates after the initial mouse vaccination results showed that it elicited the strongest neutralizing antibody response against SARS-CoV-2. Further tests in mice and hamsters indicated that PTX-COVID19-B induced robust humoral and cellular immune responses and completely protected the vaccinated animals from SARS-CoV-2 infection in the lung. Studies in hamsters also showed that PTX-COVID19-B protected the upper respiratory tract from SARS-CoV-2 infection. Mouse immune sera elicited by PTX-COVID19-B vaccination were able to neutralize SARS-CoV-2 variants of concern (VOCs), including the B.1. 1.7, B.1. 351 and P.1 lineages. No adverse effects were induced by PTX-COVID19-B in both mice and hamsters. These preclinical results indicate that PTX-COVID19-B is safe and effective. Based on these results, PTX-COVID19-B was authorized by Health Canada to enter clinical trials in December 2020 with a phase 1 clinical trial ongoing (ClinicalTrials.gov number: NCT04765436). One Sentence Summary PTX-COVID19-B is a SARS-CoV-2 mRNA vaccine that is highly immunogenic, safe, and effective in preventing SARS-CoV-2 infection in mice and hamsters and is currently being evaluated in human clinical trials.
A fluorescence-based genetic screen reveals diverse mechanisms silencing small RNA signaling in E. coli

Significance The ability to promptly switch genes on and off allows bacteria to adapt rapidly to changing environments for better survival. Many small regulatory RNAs (sRNAs), including RyhB, a sRNA made in response to iron starvation, are important switches in bacteria. We discovered factors that can keep the sRNA switch off by using a facile genetic screen platform. These factors include an RNA sponge and an adaptor protein for a ribonuclease, providing distinct perspectives on controlling sRNA signaling in bacteria. As key players of gene regulation in many bacteria, small regulatory RNAs (sRNAs) associated with the RNA chaperone Hfq shape numerous phenotypic traits, including metabolism, stress response and adaptation, as well as virulence. sRNAs can alter target messenger RNA (mRNA) translation and stability via base pairing. sRNA synthesis is generally under tight transcriptional regulation, but other levels of regulation of sRNA signaling are less well understood. Here we used a fluorescence-based functional screen to identify regulators that can quench sRNA signaling of the iron-responsive sRNA RyhB in Escherichia coli. The identified regulators fell into two classes, general regulators (affecting signaling by many sRNAs) and RyhB-specific regulators; we focused on the specific ones here. General regulators include three Hfq-interacting sRNAs, CyaR, ChiX, and McaS, previously found to act through Hfq competition, RNase T, a 3′ to 5′ exonuclease not previously implicated in sRNA degradation, and YhbS, a putative GCN5-related N-acetyltransferase (GNAT). Two specific regulators were identified. AspX, a 3′end-derived small RNA, specifically represses RyhB signaling via an RNA sponging mechanism. YicC, a previously uncharacterized but widely conserved protein, triggers rapid RyhB degradation via collaboration with the exoribonuclease PNPase. These findings greatly expand our knowledge of regulation of bacterial sRNA signaling and suggest complex regulatory networks for controlling iron homeostasis in bacteria. The fluorescence-based genetic screen system described here is a powerful tool expected to accelerate the discovery of novel regulators of sRNA signaling in many bacteria.
Scale-free behavioral dynamics directly linked with scale-free cortical dynamics

Naturally occurring body movements and collective neural activity both exhibit complex dynamics, often with scale-free, fractal spatiotemporal structure. Scale-free dynamics of both brain and behavior are each associated with functional benefits to the organism. Despite their similarities, scale-free brain activity and scale-free behavior have been studied separately, without a unified explanation. Here we show that scale-free dynamics of behavior and certain subsets of cortical neurons are strongly related. Surprisingly, the scale-free neural subsets exhibit stochastic winner-take-all competition with other neural subsets. This observation is inconsistent with current theory of critical phenomena in neural systems. We develop a computational model which incorporates known cell-type-specific circuit structure, explaining our findings with a new type of criticality. Our results establish neural underpinnings of scale-free behavior and clear behavioral relevance of scale-free neural activity. Popular Summary Over the past 2 decades many measurements of brain activity have revealed a peculiar property; the activity is statistically similar when examined at different scales. This self-similar, fractal nature of brain activity has attracted the interest of neuroscientists and physicists alike. In physics, fractal structure shows up in certain non-living systems when they are operating near the tipping point of a phase transition (e.g. liquid water turning into gaseous vapor). This led to the criticality hypothesis, which posits that the brain may also operate near the tipping point of a phase transition, i.e. near criticality. Initially the criticality hypothesis was very exciting at a basic level - physical principles of phase transitions promised to help us understand how the brain works. However, a important question naturally arose and remains unresolved. What do the scale-free, fractal fluctuations of brain activity have to do with the behavior of the organism? Is it just background noise in the brain or is there something fractal-like about how we behave and interact with the world? Here we show for the first time that scale-free fluctuations of brain activity are directly related to moment-to-moment fluctuations in behavior. In fact, we show that both the brain activity and mouse behavior have very similar fractal structure. These results take the criticality hypothesis to a new level of biological and, perhaps, psychological relevance. Scale-free brain dynamics are not just background noise; they correspond to the dynamics of behavior.
Composition and Biophysical Properties of the Sorting Platform Pods in the Shigella Type III Secretion System

Shigella flexneri, causative agent of bacillary dysentery (shigellosis), uses a type III secretion system (T3SS) as its primary virulence factor. The T3SS injectisome delivers effector proteins into host cells to promote entry and create an important intracellular niche. The injectisome’s cytoplasmic sorting platform (SP) is a critical assembly that contributes to substrate selection and energizing secretion. The SP consists of oligomeric Spa33 “pods” that associate with the basal body via MxiK and connect to the Spa47 ATPase via MxiN. The pods contain heterotrimers of Spa33 with one full-length copy associated with two copies of a C-terminal domain (Spa33C). The structure of Spa33C is known, but the precise makeup and structure of the pods in situ remains elusive. We show here that recombinant wild-type Spa33 can be prepared as a heterotrimer that forms distinct stable complexes with MxiK and MxiN. In two-hybrid analyses, association of the Spa33 complex with these proteins occurs via the full-length Spa33 component. Furthermore, these complexes each have distinct biophysical properties. Based on these properties, new high-resolution cryo-electron tomography data and architectural similarities between the Spa33 and flagellar FliM-FliN complexes, we provide a preliminary model of the Spa33 heterotrimers within the SP pods. From these findings and evolving models of SP interfaces and dynamics in the Yersinia and Salmonella T3SS, we suggest a model for SP function in which two distinct complexes come together within the context of the SP to contribute to form the complete pod structures during the recruitment of T3SS secretion substrates.
Complex introgression among three diverged largemouth bass lineages

Hybrid zones between diverged lineages offer an unique opportunity to study evolutionary processes related to speciation. Natural and anthropogenic hybridization in the black basses (Micropterus spp.) is well documented, including an extensive intergrade zone between the widespread northern Largemouth Bass (M. salmoides) and the Florida Bass (M. floridanus). Phenotypic surveys have identified an estuarine population of Largemouth Bass (M. salmoides) in the Mobile-Tensaw Delta, with larger relative weight and smaller adult size compared to inland populations, suggesting a potential third lineage of largemouth bass. To determine the evolutionary relationships between these Mobile Delta bass populations, M. salmoides, and M. floridanus, putative pure and intergrade populations of all three groups were sampled across the eastern United States. Phylogenetic analyses of 8,582 nuclear SNPs derived from genotype-by-sequencing and the ND2 mitochondrial gene determined that Delta bass populations stem from a recently diverged lineage of Largemouth Bass. Using a novel quantitative pipeline, a panel of 73 diagnostic SNPs was developed for the three lineages, evaluated for accuracy, and then used to screen 881 samples from 52 sites for genetic integrity and hybridization on the Agena MassARRAY platform. These results strongly support a redrawing of native ranges for both the intergrade zone and M. floridanus, which has significant implications for current fisheries management. Furthermore, Delta bass ancestry was shown to contribute significantly to the previously described intergrade zone between northern Largemouth Bass and Florida Bass, suggesting a more complex pattern of secondary contact and introgression among these diverged Micropterus lineages.
A rapid method to quantify small-scale vegetation patch structure to complement conventional quadrat surveys

Aims Vegetation sampling typically involves the use of quadrats, often 1m2, to estimate species cover-abundance. Such surveys do not generally record small-scale vegetation patch structure at sub-quadrat scales, for example 10 cm2. Here we test a simple method to quantify patch structure that complements conventional techniques. We compare the two methods, and analyse metrics derived from small-scale patch surveys with environment / management data. Location Northumberland, United Kingdom Methods We recorded cover-abundance of all species in an upland moor with 1m2 quadrats. These were divided into 100 ‘sub-quadrats’, 10 × 10 cm, and the dominant and subdominant species identified. Patch metrics (number, area and shape) for individual species recorded as dominant or subdominant in the sub-quadrat survey were analysed using multivariate generalised linear models with environmental and management data. Sub-quadrat data were also aggregated for each quadrat, to create species composition data. The two sets of compositional data, from whole-quadrat and sub-quadrat aggregations, were compared via Procrustes rotation of ordination scores. Results Patch number, area and shape for dominant and subdominant species were all significantly affected by soil pH, soil water content, slope and elevation. Effects of proximity to sheep tracks and drainage ditches were less consistent amongst species. Ordinations of vegetation data from conventional and sub-quadrats were similar, with significant Procrustes R-squared of 67% and 70% for dominant and subdominant species respectively. Conclusions Sub-quadrat surveys can easily be used to complement existing whole-quadrat surveys at little cost in time or resources. Their patch metrics can provide additional insights into the environmental and management drivers that may affect the growth of individual plants or clumps, potentially in relation to plant traits, and thus alter the overall community composition. The methods we describe can readily be adapted to other sizes of quadrats and sub-quadrats in a wide range of vegetation communities.
Dual functions of ZmGI1 in the photoperiodic flowering pathway and salt stress responses in maize

The circadian clock perceives photoperiodic changes and initiates processes leading to floral transition. GIGANTEA (GI) primarily functions as a principal clock component that integrates environmental cues into regulation of growth and development in Arabidopsis. However, it is unclear whether ZmGIs regulate photoperiodic flowering and abiotic stress response. Here, we demonstrated that the expression of ZmGI1 depicted a typical circadian pattern and was differentially expressed under LDs and SDs in photoperiodic sensitive and insensitive maize lines. The transcription level was significantly and positively correlated with days to silking and photoperiodic sensitivity in maize. Moreover, natural variation in ZmGI1 was associated with maize photoperiod response and the fine-tuning of plant development traits. Overexpression of ZmGI1Huangzao4 induced early flowering and enhanced salt tolerance in Arabidopsis relative to the wild-type and gi mutants. ZmGI1 formed a protein complex with ZmFKF1 and acted as a positive regulator of flowering time by regulating CONSTANS transcription in the photoperiod pathway. The ZmGI1/ZmThox complex regulates oxidative stress induced by salt stress via a redox balance pathway. Over all, we have provided compelling evidence to suggest that ZmGI1 is a pleotropic gene whose expression depicts a typical circadian rhythmic pattern and regulates flowering time and confers salt stress tolerance.
Common neural bases for processing speech prosody and music: An integrated model

Do speech and music understanding share common neural mechanisms? Here, brain bioelectrical activity was recorded in healthy participants listening to music obtained by digitally transforming speech into viola music. Sentences originally had a positive or negative affective prosody. The aim was to investigate if the emotional content of music was processed similarly to the affective prosody of speech. EEG was recorded from 128 electrodes in 20 healthy students. Participants had to detect rare neutral piano sounds while ignoring viola melodies. Negative affective valence of stimuli increased the amplitude of frontal P300 and N400 components of ERPs, while positive valence enhanced a late inferior frontal positivity. Similar markers were previously found for the processing of positive versus negative music, vocalizations, and speech. Source reconstruction showed that negative music activated the right superior temporal gyrus and cingulate cortex, while positive music activated the left middle and inferior temporal gyrus and the inferior frontal cortex. An integrated model is proposed of a possible common network for processing the emotional content of music, vocalizations, and speech, which might explain some universal and relatively innate brain reaction to music.
Mechanistic of Rac and Cdc42 synchronization at the cell edge by ARHGAP39-dependent signaling nodules and the impact on protrusion dynamics

Compartmentalization of GTPase regulators into signaling nodules dictates the GTPase pathways selected. Rac and Cdc42 are synchronized at the cell edge for effective protrusion in motile cells but how their activity is coordinated remains elusive. Here, we discovered that ARHGAP39, a Rac and Cdc42 GTPase-activating protein, sequentially interacts with WAVE and mDia2 to control Rac/lamellipodia and Cdc42/filopodia protrusions, respectively. Mechanistically, ARHGAP39 binds to WAVE and, upon phosphorylation by Src kinase, inactivates Rac to promote Cdc42-induced filopodia formation. With our optimized FRET biosensor, we detected active Cdc42 at the filopodia tips that controls filopodia extension. ARHGAP39 is transported to filopodia tips by Myosin-X where it binds mDia2 and inactivates Cdc42 leading to filopodia retraction. Failure in lamellipodia to filopodia switch by defective ARHGAP39 impairs cell invasion and metastasis. Our study reveals that compartmentalization of ARHGAP39 within Rac/Cdc42 signaling nodules orchestrates the synchronization of lamellipodia/filopodia crosstalk and highlights the intricate regulation of leading edge dynamics in migrating cells.
Remote spatially variant debiased profiling of cell and tissue mechanical properties

The role of the mechanical environment in defining tissue function, development and growth has been shown to be fundamental. Assessment of the changes in stiffness of tissue matrices at multiple scales has relied mostly on invasive and often specialist equipment such as AFM or mechanical testing devices poorly suited to the cell culture workflow.In this paper, we have developed a novel unbiased passive optical coherence elastography method, exploiting ambient vibrations in the sample that enables real-time noninvasive quantitative profiling of cells and tissues. We demonstrate a robust method that decouples optical scattering and mechanical properties by actively compensating for scattering associated noise bias and reducing variance. The efficiency for the method to retrieve ground truth is validated in silico and in vitro, and exemplified for key applications such as time course mechanical profiling of bone and cartilage spheroids, tissue engineering cancer models, tissue repair models and single cell. Our method is readily implementable with any commercial optical coherence tomography system without any hardware modifications, and thus offers a breakthrough in tissue mechanical assessment for novel on line assessment of spatial mechanical properties for organoids, soft tissues and tissue engineering.
Segregation and integration of the functional connectome in neurodevelopmentally ‘at risk’ children

Functional connectivity within and between Intrinsic Connectivity Networks (ICNs) transforms over development and supports high order cognitive functions. But how variable is this process, and does it diverge with altered cognitive developmental trajectories? We investigated age-related changes in integration and segregation within and between ICNs in neurodevelopmentally ‘at-risk’ children, identified by practitioners as experiencing cognitive difficulties in attention, learning, language, or memory. In our analysis we used performance on a battery of 10 cognitive tasks, alongside resting-state functional Magnetic Resonance Imaging in 175 at-risk children and 62 comparison children aged 5-16. We observed significant age-by-group interactions in functional connectivity between two network pairs. Integration between the ventral attention and visual networks and segregation of the limbic and fronto-parietal networks increased with age in our comparison sample, relative to at-risk children. Furthermore, functional connectivity between the ventral attention and visual networks in comparison children significantly mediated age-related improvements in executive function, compared to at-risk children. We conclude that integration between ICNs show divergent neurodevelopmental trends in the broad population of children experiencing cognitive difficulties, and that these differences in functional brain organisation may partly explain the pervasive cognitive difficulties within this group over childhood and adolescence. Research Highlights We investigated functional brain organisation and its development in 175 children who experience neurodevelopmental difficulties in cognition and behaviour, relative to a comparison sample (n=62) We replicated common neurodevelopmental trends across the samples: functional connectivity increased within Intrinsic Connectivity Networks and the default-mode network increasingly segregated with age Neurodevelopmentally at-risk children also showed different age-related changes in functional connectivity between the ventral attention and visual networks and between the fronto-parietal and limbic networks Furthermore, the integration between the ventral attention and visual networks in comparison children mediated age-related changes in cognition, relative to at-risk children