Miracle-dz/UniFAR-FTUs huggingface dataset:
Gaussian accelerated molecular dynamics reveals that a proline-rich signaling peptide frequently samples cis conformations when unbound

Disordered proline-rich motifs are common across the proteomes of many species and are often involved in protein-protein interactions. Proline is a unique amino acid due to the covalent bond between the backbone nitrogen and the proline side chain. The resulting five-membered ring allows proline to sample the cis state about its peptide bond, which other residues cannot do as readily. Because proline-rich disordered sequences exist as ensembles that likely include structures with the proline peptide bond in cis, a robust methodology to accurately account for these conformations in the overall ensemble is crucial. Observing the cis conformations of proline in a disordered sequence is challenging both experimentally and computationally. Nitrogen-hydrogen NMR spectroscopy cannot directly observe proline residues, which lack an amide bond, and computational methods struggle to overcome the large kinetic barrier between the cis and trans states, since isomerization usually occurs on the order of seconds. In the current work, Gaussian accelerated molecular dynamics was used to overcome this free energy barrier and simulate proline isomerization in a tetrapeptide (KPTP) and in the 12-residue proline-rich SH3 binding peptide, ArkA. We found that Gaussian accelerated molecular dynamics, when combined with a lowered peptide bond dihedral angle potential energy barrier (15 kcal/mol), allowed sufficient sampling of the proline cis and trans states on a microsecond timescale. All ArkA prolines spend a significant fraction of time in cis, leading to a more compact ensemble with less polyproline II helix structure than an ArkA ensemble with all peptide bonds in trans. The ensemble containing cis prolines also matches more closely to in vitro circular dichroism data than the all-trans ensemble. The ability of the ArkA prolines to isomerize likely affects the peptide’s ability to bind its partner SH3 domain, and should be studied further. This is the first molecular dynamics simulation study of proline isomerization in a biologically relevant proline-rich sequence that we know of, and a similar protocol could be applied to study multi-proline isomerization in other proline-containing proteins to improve conformational diversity and agreement with in vitro data.
Identifying components that vary in space and time from resting-state functional MRI

A widespread assumption of fMRI-derived large-scale intrinsic connectivity networks (ICNs) is that they are spatially static over time. However, the assumption of spatial stationarity of ICNs has been challenged by a range of techniques that allow for time-varying connectivity between brain regions and demonstration that canonical networks like the default model network (DMN) can be fractionated according to time-varying connectivity relationships of their subcomponents. Previously, we developed a simple spatiotemporal ICA (stICA) technique to allow the discovery of patterns of spatiotemporal evolution in task fMRI data in a way that avoided the traditional constraint of spatial stationarity on brain networks, and we validated the approach in fMRI of task-to-rest transitions. Here, we apply our stICA technique to resting-state fMRI datasets to explore whether spatiotemporally evolving components of brain activity can be identified in the absence of an overt behavioural task. We found that stICA components could generally be described in terms of graded onsets and offsets of ICNs that had been calculated based on techniques that assumed spatial stationarity. Our results suggest that, to a reasonable approximation, stable ICNs can be taken to be building blocks of the spatiotemporal patterns measured with resting-state fMRI.
Dissociation of β2m from MHC Class I Triggers Formation of Noncovalent, Transient Heavy Chain Dimers

At the plasma membrane of mammalian cells, major histocompatibility complex class I molecules (MHC-I) present antigenic peptides to cytotoxic T cells. Following the loss of the peptide and the light chain beta-2 microglobulin (β2m), the resulting free heavy chains (FHCs) can associate into homotypic complexes in the plasma membrane. Here, we investigate the stoichiometry and dynamics of MHC-I FHCs assemblies by combining a micropattern assay with fluorescence recovery after photobleaching (FRAP) and with single molecule co-tracking. We identify non-covalent MHC-I FHC dimers mediated by the α3 domain as the prevalent species at the plasma membrane, leading a moderate decrease in the diffusion coefficient. MHC-I FHC dimers show increased tendency to cluster into higher order oligomers as concluded from an increased immobile fraction with higher single molecule co-localization. In vitro studies with isolated proteins in conjunction with molecular docking and dynamics simulations suggest that in the complexes, the α3 domain of one FHC binds to another FHC in a manner similar to the β2m light chain. Significance Statement MHC class I molecules are cell surface transmembrane proteins with key functions in adaptive immunity against viral infections. The spatiotemporal organization of fully assembled MHC I at the cell surface and its function with respect to trans-interactions with T and NK cells has been studied in detail. By contrast, the consequences of peptide and β2m dissociation yielding to formation of free heavy chains (FHC) have remained unclear. We have discovered that class I free heavy chains form distinct non-covalent dimers at the cell surface rather than non-specific clustering, and we have identified a dimerization interface mediated by the α3 domain. We propose that these non-covalent dimers are the basis of distinct signaling and endocytic sorting of MHC I FHC. This is to be explored in further work.
On the contingent nature of satellite DNA evolution

Background The full catalogue of satellite DNA (satDNA) within a same genome constitutes the satellitome. The Library Hypothesis predicts that satDNA in relative species reflects that in their common ancestor, but the evolutionary mechanisms and pathways of satDNA evolution have never been analyzed for full satellitomes. We compare here the satellitomes of two Oedipodine grasshoppers (Locusta migratoria and Oedaleus decorus) which shared their most recent common ancestor about 22.8 Ma ago. Results We found that about one-third of their satDNA families (near 60 in every species) showed sequence homology, and were grouped into 12 orthologous superfamilies. The turnover rate of consensus sequences was extremely variable among the 20 orthologous family pairs analyzed in both species. The satDNAs shared by both species showed poor association with sequence signatures and motives frequently argued as functional, except for short inverted repeats allowing short dyad symmetries and non-B DNA conformations. Orthologous satDNAs frequently showed different FISH pattern at both intra- and interspecific levels. We defined indices of homogenization and degeneration, and quantified the level of incomplete library sorting between species. Conclusions Our analyses revealed that satDNA degenerates through point mutation and rejuvenates through partial turnovers caused by massive tandem duplications (the so-called satDNA amplification). Remarkably, satDNA amplification increases homogenization, at intragenomic level, and diversification between species, thus constituting the basis for concerted evolution. We suggest a model of satDNA evolution by means of recursive cycles of amplification, degeneration, and rejuvenation, leading to mostly contingent evolutionary pathways where concerted evolution emerges promptly after lineages split.
Behavioral and fMRI evidence that arousal enhances bottom-up attention and memory selectivity in young but not older adults

During a challenge or emotional experience, increases in arousal help us focus on the most salient or relevant details and ignore distracting stimuli. The noradrenergic system integrates signals about arousal states throughout the brain and helps coordinate this adaptive attentional selectivity. However, age-related changes in the noradrenergic system and attention networks in the brain may reduce the efficiency of arousal to modulate selective processing in older adults. In the current neuroimaging study, we examined age differences in how arousal affects bottom-up attention to category-selective stimuli differing in perceptual salience. We found a dissociation in how arousal modulates selective processing in the young and older brain. In young adults, emotionally arousing sounds enhanced selective incidental memory and brain activity in the extrastriate body area for salient versus non-salient images of bodies. Older adults showed no such advantage in selective processing under arousal. These age differences could not be attributed to changes in the arousal response or less neural distinctiveness in old age. Rather, our results suggest that, relative to young adults, older adults become less effective at focusing on salient over non-salient details during increases in emotional arousal.
Highly Efficient Synthetic CRISPR RNA/Cas9-Based Mutagenesis for Rapid Cardiovascular Phenotypic Screening in F0 Zebrafish

The zebrafish is a valuable vertebrate model to study cardiovascular formation and function due to the facile visualization and rapid development of the circulatory system in its externally growing embryos. Despite having distinct advantages, zebrafish have paralogs of many important genes, making reverse genetics approaches inefficient since generating animals bearing multiple gene mutations requires substantial efforts. Here, we present a simple and robust synthetic CRISPR RNA/Cas9-based mutagenesis approach for generating biallelic F0 zebrafish knockouts. Using a dual-guide synthetic CRISPR RNA/Cas9 ribonucleoprotein (dgRNP) system, we compared the efficiency of biallelic gene disruptions following the injections of one, two, and three dgRNPs per gene into the cytoplasm or yolk. We show that simultaneous cytoplasmic injections of three distinct dgRNPs per gene into one-cell stage embryos resulted in the most efficient and consistent biallelic gene disruptions. Importantly, this triple dgRNP approach enables efficient inactivation of cell autonomous and cell non-autonomous gene function, likely due to the low mosaicism of biallelic disruptions. In support of this finding, we provide evidence that the F0 animals generated by this method fully phenocopied the endothelial and peri-vascular defects observed in corresponding stable mutant homozygotes. Moreover, this approach faithfully recapitulated the trunk vessel phenotypes resulting from the genetic interaction between two vegfr2 zebrafish paralogs. Mechanistically, investigation of genome editing and mRNA decay indicates that the combined mutagenic actions of three dgRNPs per gene lead to an increased probability of frameshift mutations, enabling efficient biallelic gene disruptions. Therefore, our approach offers a highly robust genetic platform to quickly assess novel and redundant gene function in F0 zebrafish.
Oil Palm Contract Farming in Brazil: Labour Constraints and Inclusivity Challenges

Abstract The Government of Brazil established their Sustainable Palm Oil Production Programme (SPOPP) in 2010, which sought to enhance the sustainability and inclusiveness of oil palm development in the Amazon. This paper evaluates how well oil palm contract farming promoted by SPOPP has delivered on its inclusive development objectives. Drawing on cross-section data collected in Northeast Pará, it analyses two recurrent SPOPP themes, namely (1) equitable participation and (2) labour allocation to plantation management. Our analysis demonstrates that household availability of land and labour resources strongly shapes patterns of inclusion and exclusion. Moreover, findings reveal that labour time allocation is influenced by hiring of external labourers, which increases when households are labour and land poor. These results give reason to question the utility of labour-oriented contract farming eligibility criteria, revealing important inclusive business and value chain development dilemmas.
Meglumine catalyzed one pot synthesis of new fluorescent 2-amino-4-pyrazolyl-6-aryldiazenyl-4H-chromene-3-carbonitriles

Abstract Meglumine, a biodegradable basic organo-catalyst has been efficiently explored for the one-pot synthesis of new fluorescent 2-amino-4-(5-hydroxy-3-methyl-1H-pyrazol-4-yl)-6-aryldiazenyl-4H-chromene-3-carbonitriles in an aqueous medium at room temperature. The synthesized compounds were found highly fluorescent when screened for photoluminescence properties. The planar structure equipped with substituted aryldiazenyl group led to the extension of conjugation that facilitated fluorescence emission in the visible region with a large stokes shift of 290–294 nm. The novelty of work is a synthesis of highly conjugated molecular assembly, high yield in shorter reaction time, energy efficiency, atom economy, utilization of water as a universal green solvent and meglumine as an eco-benign organo-catalyst. Graphical Abstract
Introduction to the special issue on Volcanism in Zealandia and the SW Pacific

ABSTRACT The New Zealand Journal of Geology and Geophysics has prepared a two-part Special Issue on volcanism in Zealandia and the southwest Pacific. Part 1, published in 2020 as Issue 4 of Volume 63, summarises regional aspects of Zealandia volcanism as well as South Island and Antarctica volcanism. Part 2, published in 2021 as joint issue 2–3 of Volume 64, summarises North Island volcanism. In this preface, I summarise the content of these issues and point out some (but certainly not all) outstanding topics.
Microwave-Assisted Synthesis of Carbon Dot – Iron Oxide Nanoparticles for Fluorescence Imaging and Therapy

Fluorescence microscopy is commonly used to image specific parts of a biological system, and is applicable for early diagnosis of cancer. Current fluorescent probes, such as organic dyes and quantum dots, suffer from poor solubility and high toxicity, respectively, demonstrating a need for a colloidal stable and non-toxic fluorescent probe. Here we present an iron oxide and carbon dot (CD) based nanoparticle (CNPCP) that displays optical properties similar to those of conventional fluorescent probe and also exhibits good biocompatibility. Fluorescent CDs were synthesized from glucosamine onto chitosan – polyethylene glycol (PEG) graft copolymer using microwave irradiation. These NPs were monodispersed in aqueous environments and displayed excitation-dependent fluorescence; they demonstrated good size stability and fluorescence intensity in biological media. In vitro evaluation of CNP as fluorescent probes in cancer cell lines showed that these NPs caused little toxicity, and allowed fast and quantitative imaging. Model therapeutic doxorubicin (DOX) was conjugated onto the NPs (CNPCP-DOX) to demonstrate the multifunctionality of the NPs, and in vitro studies showed that CNPCP-DOX was able to kill cancer cells in a dose dependent manner. These results indicate the potential of using CNPCPs as fluorescent probes capable of delivering chemotherapeutics.
Genome-Wide Identification of the Early Flowering 4 (ELF4) Gene Family in Cotton and Silent GhELF4-1 and GhEFL3-6 Decreased Cotton Stress Resistance

The early flowering 4 (ELF4) family members play multiple roles in the physiological development of plants. ELF4s participated in the plant biological clock’s regulation process, photoperiod, hypocotyl elongation, and flowering time. However, the function in the ELF4s gene is barely known. In this study, 11, 12, 21, and 22 ELF4 genes were identified from the genomes of Gossypium arboreum, Gossypium raimondii, Gossypium hirsutum, and Gossypium barbadense, respectively. There ELF4s genes were classified into four subfamilies, and members from the same subfamily show relatively conservative gene structures. The results of gene chromosome location and gene duplication revealed that segmental duplication promotes gene expansion, and the Ka/Ks indicated that the ELF4 gene family has undergone purification selection during long-term evolution. Spatio-temporal expression patterns and qRT-PCR showed that GhELF4 genes were mainly related to flower, leaf, and fiber development. Cis-acting elements analysis and qRT-PCR showed that GhELF4 genes might be involved in the regulation of abscisic acid (ABA) or light pathways. Silencing of GhELF4-1 and GhEFL3-6 significantly affected the height of cotton seedlings and reduced the resistance of cotton. The identification and functional analysis of ELF4 genes in upland cotton provide more candidate genes for genetic modification.
Biosynthesis of Fusapyrone Depends on the H3K9 Methyltransferase, FmKmt1, in Fusarium mangiferae

The phytopathogenic fungus Fusarium mangiferae belongs to the Fusarium fujikuroi species complex (FFSC). Members of this group cause a wide spectrum of devastating diseases on diverse agricultural crops. F. mangiferae is the causal agent of the mango malformation disease (MMD) and as such detrimental for agriculture in the southern hemisphere. During plant infection, the fungus produces a plethora of bioactive secondary metabolites (SMs), which most often lead to severe adverse defects on plants health. Changes in chromatin structure achieved by posttranslational modifications (PTM) of histones play a key role in regulation of fungal SM biosynthesis. Posttranslational tri-methylation of histone 3 lysine 9 (H3K9me3) is considered a hallmark of heterochromatin and established by the SET-domain protein Kmt1. Here, we show that FmKmt1 is involved in H3K9me3 in F. mangiferae. Loss of FmKmt1 only slightly though significantly affected fungal hyphal growth and stress response and is required for wild type-like conidiation. While FmKmt1 is largely dispensable for the biosynthesis of most known SMs, removal of FmKMT1 resulted in an almost complete loss of fusapyrone and deoxyfusapyrone, γ-pyrones previously only known from Fusarium semitectum. Here, we identified the polyketide synthase (PKS) FmPKS40 to be involved in fusapyrone biosynthesis, delineate putative cluster borders by co-expression studies and provide insights into its regulation.
Detecting Roles of Money Laundering in Bitcoin Mixing Transactions: A Goal Modeling and Mining Framework

Cryptocurrency has become a new venue for money laundering. Bitcoin mixing services deliberately obfuscate the relationship between senders and recipients, making it difficult to trace suspicious money flow. We believe that the key to demystifying the bitcoin mixing services is to discover agents’ roles in the money laundering process. We propose a goal-oriented approach to modeling, discovering, and analyzing different types of roles in the agent-based business process of the bitcoin mixing scenario using historical bitcoin transaction data. It adopts the agents’ goal perspective to study the roles in the bitcoin money laundering process. Moreover, it provides a foundation to discover real-world agents’ roles in bitcoin money laundering scenarios.
Meta-Analyzing Multiple Omics Data With Robust Variable Selection

High-throughput omics data are becoming more and more popular in various areas of science. Given that many publicly available datasets address the same questions, researchers have applied meta-analysis to synthesize multiple datasets to achieve more reliable results for model estimation and prediction. Due to the high dimensionality of omics data, it is also desirable to incorporate variable selection into meta-analysis. Existing meta-analyzing variable selection methods are often sensitive to the presence of outliers, and may lead to missed detections of relevant covariates, especially for lasso-type penalties. In this paper, we develop a robust variable selection algorithm for meta-analyzing high-dimensional datasets based on logistic regression. We first search an outlier-free subset from each dataset by borrowing information across the datasets with repeatedly use of the least trimmed squared estimates for the logistic model and together with a hierarchical bi-level variable selection technique. We then refine a reweighting step to further improve the efficiency after obtaining a reliable non-outlier subset. Simulation studies and real data analysis show that our new method can provide more reliable results than the existing meta-analysis methods in the presence of outliers.
Fear Memory Retrieval Is Associated With a Reduction in AMPA Receptor Density at Thalamic to Amygdala Intercalated Cell Synapses

The amygdala plays a crucial role in attaching emotional significance to environmental cues. Its intercalated cell masses (ITC) are tight clusters of GABAergic neurons, which are distributed around the basolateral amygdala complex. Distinct ITC clusters are involved in the acquisition and extinction of conditioned fear responses. Previously, we have shown that fear memory retrieval reduces the AMPA/NMDA ratio at thalamic afferents to ITC neurons within the dorsal medio-paracapsular cluster. Here, we investigate the molecular mechanisms underlying the fear-mediated reduction in the AMPA/NMDA ratio at these synapses and, in particular, whether specific changes in the synaptic density of AMPA receptors underlie the observed change. To this aim, we used a detergent-digested freeze-fracture replica immunolabeling technique (FRIL) approach that enables to visualize the spatial distribution of intrasynaptic AMPA receptors at high resolution. AMPA receptors were detected using an antibody raised against an epitope common to all AMPA subunits. To visualize thalamic inputs, we virally transduced the posterior thalamic complex with Channelrhodopsin 2-YFP, which is anterogradely transported along axons. Using face-matched replica, we confirmed that the postsynaptic elements were ITC neurons due to their prominent expression of μ-opioid receptors. With this approach, we show that, following auditory fear conditioning in mice, the formation and retrieval of fear memory is linked to a significant reduction in the density of AMPA receptors, particularly at spine synapses formed by inputs of the posterior intralaminar thalamic and medial geniculate nuclei onto identified ITC neurons. Our study is one of the few that has directly linked the regulation of AMPA receptor trafficking to memory processes in identified neuronal networks, by showing that fear-memory induced reduction in AMPA/NMDA ratio at thalamic-ITC synapses is associated with a reduced postsynaptic AMPA receptor density.