title: The relationship between intelligence and orientation discrimination skills

abstract:
We investigate the notion that basic visual information is acting as a building block for more complex cognitive processes. Specifically, we measure individual orientation discrimination thresholds report good correlations against IQ, verbal-IQ and non-verbal IQ scores. Further we calculate correlations between the four WASI-II subtests and orientation discrimination thresholds, with Matrix Reasoning maintaining the strongest relationship, even when controlling for the effects of the other three subtests. Vocabulary raw scores which quantify the ability to describe verbally a specific word, showed the second strongest correlation, when controlling for the effects of Matrix Reasoning and Block Design, but not Similarities. Our results demonstrate that low-level visual abilities and high-level cognitive processes are more tightly interwoven together than previously thought, generating evidence to show that vision is tightly linked to human cognition.

Statement of RelevanceVision is one of the building blocks of our world experience through thoughts, emotions and actions. It is undeniably the sense upon which we rely the most, while working, driving, reading a book, dancing. So, it should come as no surprise that the intelligence quotient (IQ), widely used in the modern world to assess the cognitive abilities of a person in relation to their age group correlates well with visual abilities. Previous studies have demonstrated that pitch and colour discrimination, motion discrimination and contrast discrimination abilities are tightly linked to IQ. In this study we demonstrate for the first time, that the simplest visual ability, orientation discrimination which is computed by neurons in the primary visual cortex is tightly linked to intelligence. We believe a simple and quick orientation discrimination task could be used as an index of intelligence in clinical populations and other species, in addition to traditional measures.


title: Gene expression in Lucilia sericata (Diptera: Calliphoridae) larvae exposed to Pseudomonas aeruginosa and Acinetobacter baumanii identifies shared and microbe-specific induction of immune genes

abstract: Antibiotic resistance is a continuing challenge in medicine. There are various strategies for expanding antibiotic therapeutic repertoires, including the use of blow flies. Their larvae exhibit strong antibiotic and antibiofilm properties that alter microbiome communities. One species, Lucilia sericata, is used to treat problematic wounds due to its debridement capabilities and its excretions and secretions that kill some pathogenic bacteria. There is much to be learned about how L. sericata interacts with microbiomes at the molecular level. To address this deficiency, gene expression was assessed after feeding exposure (1 hour or 4 hours) to two clinically problematic pathogens: Pseudomonas aeruginosa and Acinetobacter baumanii. The results identified immunity related genes that were differentially expressed when exposed to these pathogens, as well as non-immune genes possibly involved in gut responses to bacterial infection. There was a greater response to P. aeruginosa that increased over time, while few genes responded to A. baumanii exposure and expression was not time-dependent. The response to feeding on pathogens indicates a few common responses and features distinct to each pathogen, which is useful in improving wound debridement therapy and helps develop biomimetic alternatives.

category: immunology


title: The response of desert biocrust bacterial communities to hydration-desiccation cycles

abstract: Rain events in arid environments are highly unpredictable, interspersing extended periods of drought. Therefore, following changes in desert soil bacterial communities during hydration-desiccation cycles in the field, was seldom attempted. Here, we assessed rain-mediated dynamics of active community in the Negev Desert biological soil crust (biocrust), and evaluated the changes in bacterial composition, potential function, and photosynthetic activity. We predicted that increased biocrust moisture would resuscitate the phototrophs, while desiccation would inhibit their activity. Our results show that hydration increased chlorophyll content, resuscitated the biocrust Cyanobacteria, and induced potential phototrophic functions. However, decrease in the soil water content did not immediately decrease the phototrophs activity, though chlorophyll levels decreased. Moreover, while the Cyanobacteria relative abundance significantly increased, Actinobacteria, the former dominant taxa, significantly decreased in abundance. We propose that, following a rain event biocrust moisture significantly decreased, almost to drought levels, yet the response of the active bacterial community lagged, in contrast to topsoil. Possible explanations to the described rain-mediated bacteria dynamics are discussed.

category: microbiology


title: Halfway to self-sustainability: Reintroduced migratory European Northern Bald Ibises (Geronticus eremita) still need management interventions for population viability

abstract:
Northern Bald Ibis (NBI) have disappeared from Europe already in Middle Age. Since 2003 a migratory population is reintroduced in Central Europe. We conducted demographic analyses of survival and reproduction of 384 NBI over a period of 12 years (2008-2019). These data also formed the basis for a population viability analysis (PVA) simulating the possible future development of the NBI population in different scenarios. We tested life-stage specific survival rates for differences between these stages, raising types and colonies as well as the influence of stochastic events and NBI supplements on the population growth.

Stage specific survival rates ranged from 0.64 to 0.78. 61% of the mature females reproduce with a mean fecundity of 2.15 fledglings per nest. The complementary PVA indicated that the release population is close to self-sustainability with a given lambda 0.95 and 24% extinction probability within 50 years. Of the 326 future scenarios tested, 94 % reached the criteria of <5% extinction probability and population growth rates >1. In case of positive population growth, stochastic events had a limited effect. Of 820 sub-scenarios with different stochastic event frequencies and severities 87 % show population growth despite the occurrence of stochastic events.

Predictions can be made based on the results of the individual-based model as to whether and under what circumstances the reintroduced NBI population can survive. This study shows that a PVA can support reintroduction success that should work closely together with the project in the field for mutual benefit, to optimize future management decisions.

category: ecology


title: An extended admixture pulse model reveals the limits to the dating of Human-Neandertal introgression

abstract: Neandertal DNA makes up 2-3% of the genomes of all non-African individuals. The patterns of Neandertal ancestry in modern humans have been used to estimate that this is the result of gene flow that occurred during the expansion of modern humans into Eurasia, but the precise dates of this event remain largely unknown. Here, we introduce an extended admixture pulse model that allows joint estimation of the timing and duration of gene flow. This model leads to simple expressions for both the admixture segment distribution and the decay curve of ancestry linkage disequilibrium, and we show that these two statistics are closely related. In simulations, we find that estimates of the mean time of admixture are largely robust to details in gene flow models, but that the duration of the gene flow can only be recovered if gene flow is very recent and the exact recombination map is known. These results imply that gene flow from Neandertals into modern humans could have happened over hundreds of generations. Ancient genomes from the time around the admixture event are thus likely required to resolve the question when, where, and for how long humans and Neandertals interacted.

category: evolutionary biology


title: An extended admixture pulse model reveals the limitations to Human-Neandertal introgression dating


title: Genetic diversity in facultatively sexual populations and its implications for the origins of self-incompatibility in algae and fungi

abstract:
The evolutionary mechanism that drove the establishment of self-incompatibility in early sexual eukaryotes is still a debated topic. While a number of competing hypotheses have been proposed, many have not received detailed theoretical attention. In particular, the hypothesis that self-incompatibility increases the benefits of genetic recombination in sexual haploids has been comparatively understudied. In this paper we address this topic by mathematically deriving how the probability of mating with a genetically distinct individual changes as a function of the presence or absence of self-incompatible mating type classes. We find that although populations with mating types successfully engage in sexual reproduction less frequently than their self-compatible competitors, they can nevertheless engage in useful sex with genetically distinct partners more frequently. This conclusion holds when the number of sexual reproductive events per generation is low (i.e. in small populations with low rates of facultative sexual reproduction). Finally we demonstrate the potential for frequency-dependent selection in competitive dynamics between self-compatible and self-incompatible types. These analytic results provide a baseline for studying the sex advantage enhancer model for the evolutionary origin of mating types within each specific hypothesis for the evolution of recombination.

PACS87.23.-n Ecology and evolution, 87.23.Kg Dynamics of evolution, 02.50.Ey Stochastic processes

2000 MSC37N25: Dynamical systems in biology, 60J70: Applications of diffusion theory (population genetics, absorption problems, etc.)


title: Selective functional inhibition of a tumor-derived p53 mutant by cytosolic chaperones identified using split-YFP in budding yeast

abstract: Life requires the oligomerization of individual proteins into higher-order assemblies. In order to form functional oligomers, monomers must adopt appropriate three-dimensional structures. Molecular chaperones transiently bind nascent or misfolded proteins to promote proper folding. Single missense mutations frequently cause disease by perturbing folding despite chaperone engagement. A misfolded mutant capable of oligomerizing with wild-type proteins can dominantly poison oligomer function. We previously found evidence that human-disease-linked mutations in Saccharomyces cerevisiae septin proteins slow folding and attract chaperones, resulting in a kinetic delay in oligomerization that prevents the mutant from interfering with wild-type function. Here we build upon our septin studies to develop a new approach to identifying chaperone interactions in living cells, and use it to expand our understanding of chaperone involvement, kinetic folding delays, and oligomerization in the recessive behavior of tumor-derived mutants of the tumor suppressor p53. We find evidence of increased binding of several cytosolic chaperones to a recessive, misfolding-prone mutant, p53(V272M). Similar to our septin results, chaperone overexpression inhibits the function of p53(V272M) with minimal effect on the wild type. Unlike mutant septins, p53(V272M) is not kinetically delayed under conditions in which it is functional. Instead, it interacts with wild-type p53 but this interaction is temperature sensitive. At high temperatures or upon chaperone overexpression, p53(V272M) is excluded from the nucleus and cannot function or perturb wild-type function. Chaperone inhibition liberates the mutant to enter the nucleus where it has a slight dominant-negative effect. These findings provide new insights into the effects of missense mutations.

category: genetics


title: Integrative RNA-Seq analysis of Capsicum annuum L.-Phytophthora capsici L. pathosystem reveals molecular cross-talk and activation of host defence response

abstract:
BackgroundChili pepper (Capsicum annuum L.) being one of an important member of the Solanaceae family, and its productivity is highly affected by the fungal pathogen Phytophthora capsici L. Other to CM-344, the unavailability of resistant landraces to all possible strains of P. capsici imposes a serious threat to its global production. This is because of our current understanding of the molecular mechanisms associated with the defence response in C. annuum-P. capsici pathosystem is limited. Therefore, the current study used RNA-seq technology to dissect the genes associated with defence response against P. capsici infection in two contrasting landraces, i.e. GojamMecha_9086 (Resistant) and Dabat_80045 (susceptible) exposed to P. capsici infection.

ResultsThe transcriptomes from 4 leaf samples (RC, RI, SC and SI) of chili pepper resulted in a total of 1,18, 879 assembled transcripts (with a mean TL of 813.23bp and N50 of 1,277bp) along with 52,384 pooled unigenes with (mean UL of 1029.36 bp and N50 of 1,403bp). The enrichment analysis of the transcripts indicated 23 different KEGG pathways under five main categories. Further, 774 and 484 differentially expressed genes (DEGs) were obtained from RC vs. RI and SC vs. SI leaf samples, respectively. Of these, 57 DEGs were found to be associated with defence responses against P. capsici infection. The defence-related genes, such as LTPL, defensin J1-2-like, peroxidase 5-like, UGT, and GRP proteins-like, were more significantly upregulated in RC vs. RI. Furthermore, RT-qPCR analysis of six randomly selected genes validated the results of Illumina NextSeq500 sequencing results. Furthermore, a total of 58 TF families (bHLH most abundant) and 2,095 protein families (Protein kinase, PF00069, most abundant) were observed across all the samples with maximum hits in RI and SI samples.

ConclusionsRNA-Seq analysis of chili peppers during P.capsici infection revealed differential regulation of genes associated with defence and signaling response with shared coordination of molecular function, cellular component and biological processing. The results presented here would enhance our present understanding of the defence response in chili pepper against P. capsici infection, which could be utilized by the molecular breeders to develop resistant chili genotypes.

category: genomics


title: Cobolt: Joint analysis of multimodal single-cell sequencing data

abstract: A growing number of single-cell sequencing platforms enable joint profiling of multiple omics from the same cells. We present Cobolt, a novel method that not only allows for analyzing the data from joint-modality platforms, but provides a coherent framework for the integration of multiple datasets measured on different modalities. We demonstrate its performance on multi-modality data of gene expression and chromatin accessibility and illustrate the integration abilities of Cobolt by jointly analyzing this multi-modality data with single-cell RNA-seq and ATAC-seq datasets.

category: bioinformatics


title: UnMICST: Deep learning with real augmentation for robust segmentation of highly multiplexed images of human tissues

abstract: Newly developed technologies have made it feasible to routinely collect highly multiplexed (20-60 channel) images at subcellular resolution from human tissues for research and diagnostic purposes. Extracting single cell data from such images requires efficient and accurate image segmentation, a challenging problem that has recently benefited from the use of deep learning. In this paper, we demonstrate two approaches to improving tissue segmentation that are applicable to multiple deep learning frameworks. The first uses "real augmentations" that comprise defocused and saturated image data collected on the same instruments as the actual data; using real augmentation improves model accuracy to a significantly greater degree than computational augmentation (Gaussian blurring). The second involves imaging the nuclear envelope to better identify nuclear outlines. The two approaches cumulatively and substantially improve segmentation on a wide range of tissue types and provide a set of improved segmentation models. We speculate that the use of real augmentations may have applications in image processing outside of microscopy.


title: Deuterium Magnetic Resonance Spectroscopy of Early Treatment-Induced Changes in Tumour Lactate in vitro

abstract: Elevated production of lactate is a key characteristic of aberrant tumour cell metabolism and can be non-invasively measured as an early marker of tumour response using deuterium (2H) magnetic resonance spectroscopy (MRS). Following treatment, changes in the 2H-labeled lactate signal could identify tumour cell death or impaired metabolic function, which precede morphological changes conventionally used to assess tumour response. In this work, the association between apoptotic cell death, extracellular lactate concentration, and early treatment-induced changes in the 2H-labeled lactate signal was established in an in vitro tumour model. Experiments were conducted at 7 T on acute myeloid leukemia cells which had been treated with 10 {micro}g/mL of the chemotherapeutic agent cisplatin. At 24 and 48 hours after cisplatin treatment, the cells were injected with 20 mM of [6,6-2H2]glucose and scanned over two hours using a two-dimensional 2H MR spectroscopic imaging sequence. The resulting signals from 2H-labeled glucose, lactate, and water were quantified using a spectral fitting algorithm implemented on the OXford Spectroscopy Analysis (OXSA) MATLAB toolbox. After scanning, the cells were processed for histological stains (TUNEL [terminal deoxynucleotidyl transferase UTP nick end labeling] and H&E [hematoxylin and eosin]) to assess apoptotic area fraction and cell morphology respectively, while a colorimetric assay was used to measure extracellular lactate concentrations in the supernatant. Significantly lower levels of 2H-labeled lactate were observed in the 48-hour treated cells compared to the untreated and 24-hour treated cells, and these changes were significantly correlated with an increase in apoptotic fraction and a decrease in extracellular lactate. By establishing the biological processes associated with treatment-induced changes in the 2H-labeled lactate signal, these findings suggest that 2H MRS of lactate may be valuable in evaluating early tumour response.

category: biophysics


title: Fungal jasmonate as a novel morphogenetic signal for pathogenesis

abstract: A key question that has remained unanswered is how pathogenic fungi switch from vegetative growth to infection-related morphogenesis during a disease cycle. Here, we identify a fungal oxylipin analogous to the well-known phytohormone jasmonic acid, as the principal morphogenesis signal responsible for such a developmental switch to pathogenicity in the rice-blast fungus Magnaporthe oryzae. We explored the molecular function(s) of such intrinsic jasmonic acid during pathogenic differentiation in M. oryzae via OPR1, which encodes a 12-Oxo-phytodienoic Acid Reductase essential for its biosynthesis. Loss of OPR1 led to prolonged vegetative growth, and a delayed initiation and improper development of infection structures in M. oryzae, reminiscent of phenotypes observed in mutants (e.g. pth11{Delta} and cpka{Delta}) that are compromised for cyclic AMP signaling. Genetic- or chemical-complementation completely restored proper germ tube growth and appressorium formation in opr1{Delta}. Liquid chromatography mass spectrometry-based quantification revealed increased OPDA accumulation and a significant decrease in JA levels in the opr1{Delta}. Most interestingly, exogenous jasmonic acid also restored appressorium formation in the pth11{Delta} mutant that lacks G protein/cyclic AMP signaling. Epistasis analysis placed fungal jasmonate upstream of the cyclic AMP signaling in rice blast. Lastly, we show that intrinsic jasmonate orchestrates the cessation of vegetative phase and initiates pathogenic development via a regulatory interaction with the cyclic AMP cascade and redox signaling in rice blast.

category: cell biology


title: ANP32A represses Wnt signaling across tissues tissues thereby protecting against joint and heart disease

abstract: Wnt signaling is key to diverse homeostatic and pathological processes. This cascade is hyper-activated in osteoarthritis, the most common joint disease. Yet, fundamental aspects of Wnt signaling remain undiscovered. Here, we report that ANP32A negatively regulates Wnt signaling across tissues. In cartilage, loss of Anp32a triggered Wnt hyper-activation. Mechanistically, ANP32A directly interacted with Wnt pathway components and inhibited Wnt target genes via histone acetylation masking. Wnt antagonist treatment reduced severity of osteoarthritis in Anp32a-deficient mice preventing osteophyte formation, contrasting with cartilage-protective effects of ANP32A on oxidative stress. Hence, dual therapy targeting Wnt signaling and oxidative stress in Anp32a-deficient mice ameliorated more osteoarthritis features than individual treatments. Anp32a loss also resulted in Wnt hyper-activation in the heart with cardiac hypertrophy, and in the hippocampus, shedding light on mechanisms for reported links between ANP32A and Alzheimers disease. Collectively, this work reveals that ANP32A is a translationally relevant repressor of Wnt signaling, impacting homeostasis and disease across tissues.


title: Directional reorientation of migrating neutrophils is limited by suppression of receptor input signaling at the cell rear through myosin II activity

abstract: To migrate efficiently to target locations, cells must integrate receptor inputs while maintaining polarity: a distinct front that leads and a rear that follows. Here we investigate what is necessary to overwrite pre-existing front/rear polarity in neutrophil-like HL60 cells migrating inside straight microfluidic channels. Using subcellular optogenetic receptor activation, we show that receptor inputs can reorient weakly polarized cells, but the rear of strongly polarized cells is refractory to new inputs. Transient stimulation reveals a multi-step repolarization process, confirming that cell rear sensitivity to receptor input is the primary determinant of large-scale directional reversal. We demonstrate that the RhoA/ROCK/myosin II pathway limits the ability of receptor inputs to signal to Cdc42 and reorient migrating neutrophils. We discover that by tuning the phosphorylation of myosin regulatory light chain we can modulate the activity and localization of myosin II and thus the amenability of the cell rear to  listen to receptor inputs and respond to directional reprogramming.


title: A translation control module coordinates germline stem cell differentiation with ribosome biogenesis during Drosophila oogenesis

abstract: Ribosomal defects perturb stem cell differentiation, causing diseases called ribosomopathies. How ribosome levels control stem cell differentiation is not fully known. Here, we discovered three RNA helicases are required for ribosome biogenesis and for Drosophila oogenesis. Loss of these helicases, which we named Aramis, Athos and Porthos, lead to aberrant stabilization of p53, cell cycle arrest and stalled GSC differentiation. Unexpectedly, Aramis is required for efficient translation of a cohort of mRNAs containing a 5-Terminal-Oligo-Pyrimidine (TOP)-motif, including mRNAs that encode ribosomal proteins and a conserved p53 inhibitor, Novel Nucleolar protein 1 (Non1). The TOP-motif co-regulates the translation of growth-related mRNAs in mammals. As in mammals, the La-related protein co-regulates the translation of TOP-motif containing RNAs during Drosophila oogenesis. Thus, a previously unappreciated TOP-motif in Drosophila responds to reduced ribosome biogenesis to co-regulate the translation of ribosomal proteins and a p53 repressor, thus coupling ribosome biogenesis to GSC differentiation.

category: developmental biology


title: Deacetylation of IFIT2 mediated by HDAC5 promotes the stemness and progression of glioma

abstract:
BackgroundGlioma is the most common primary brain tumor, and the tumor stemness is a major regulatory factor affecting the progression, metastasis and recurrence of glioma. Recent research has shown that, nonhistone acetylation is widely involved in key cellular processes, including stemness regulation. The deacetylase inhibitors are promising new drugs, but their application and molecular mechanism in glioma have not been elucidated.

MethodsCCK8 and colony formation assay were used to detect cell proliferation, transwell assay was used to detect cell migration, flow cytometry was used to analyze cell apoptosis, sphere formation assay and western blot were used to detect the status of stemness. RNA-sequence, quantitative PCR and western blot were performed to screen the key molecules mediating LBH589 function. Immunoprecipitation (IP) and western blot were used to analyze the acetylation level of IFIT2. The SiRNA target HDAC4 or HDAC5, overexpression plasmids of acetyltransferases were used to identify the acetyltransferase and deacetylase regulating IFIT2. The regulatory mechanism was explored by IP and ubiquitination analysis. Finally, the xenograft tumor model in nude mice was constructed and further analyzed in vivo.

ResultsThe data showed that IFIT2 mediates the HDACi LBH589 inhibition on cell proliferation, migration and stemness, and contribution to autophagy and apoptosis in glioma. And the down-regulation of IFIT2 in glioma was confirmed to be related to its deacetylation by overexpression HDAC5, which promotes the stemness and progression of glioma. Further, deacetylation of IFIT2 by HDAC5 was demonstrated to induce its ubiquitination and subsequent protein instability, which led to loss of anti-tumor activity for IFIT2, and acceleration to glioma stemness and progression. In addition, the results indicated that IFIT2 inhibits PKC pathway, and suppressing of IFIT2 promotes tumor growth in vivo.

ConclusionsThese results not only clarify a novel post-transcriptional regulatory mode of IFIT2, but also provide a new sight of molecular mechanism for HDACi in glioma.

category: cancer biology


title: From treadmill to trails: predicting performance of runners

abstract:
Previous laboratory studies have measured the energetic costs to humans of running at uphill and downhill slopes on a treadmill. This work investigates the extension of those results to the prediction of relative performance of athletes running on flat, hilly, or very mountainous outdoor courses. Publicly available race results in the Los Angeles area provided a set of 109,000 times, with 2200 runners participating in more than one race, so that their times could be compared under different conditions. I compare with the results of a traditional model in which the only parameters considered are total distance and elevation gain. Both the treadmill-based model and the gain-based model have some shortcomings, leading to the creation of a hybrid model that combines the best features of each.

Author summaryRunning a race on a road allows absolute measures of performance. Trail running, however, has traditionally been thought of as a sport in which the only valid comparison is between different runners competing on the same course on the same day. Even the exact measurement of distance is considered to be unimportant, since courses and conditions vary so much.

An extreme example is the relatively new genre of "vertical" races, in which runners race up a mountain. In a typical example, the competitors cover a horizontal distance of 5 km, while climbing about 1000 m. The winner in one such race had a time almost triple that expected for a state-champion high school runner in a 5k road race. Clearly no comparison can be made here without taking into account the amount of climbing.

In noncompetitive contexts, many runners venture onto mountain trails, lightly dressed and with little equipment, so that it becomes important to be able to anticipate whether they will have the endurance needed to be able to safely complete a planned route. Again, this is impossible without some model of the effect of hill climbing.

category: physiology


title: Case based learning as a dynamic approach towards learning oral pathology.

abstract:
IntroductionOral pathology comprises of extensive teaching-learning of histopathology. Teaching histopathology is always challenging more when numerous histopathology slides are involved. There is lack of interest among the students and also face challenges to correlate with the clinical presentation. For effective learning, need for a method which is interactive and helps create clinico pathological correlation was felt. This study was designed to assess the impact and feasibility of case-based learning in routine teaching among dental students.

MethodsA cross sectional study was conducted among 58 undergraduate dental students, wherein a case-based learning method was introduced in the practical classes. Thirty students were randomly selected as case based learning and remaining as conventional group. Multiple paper-based cases on oral cancer was designed and used for the case based learning group of students. A self-designed pre and posttest interventional tools along with post intervention assessment using modified essay question was designed and applied.

ResultsSignificant difference in the scores between pre and post intervention questionnaire was observed among the case-based learning group (p<0.0001), however non-significant among the conventional. Similarly, significant difference in the scores of modified essay question was observed between the study groups. Majority of the students among the case-based learning group agreed that it benefitted their personal, professional and communicative skills. The students expressed their enthusiasm in learning with this method and suggested to apply it more often.

DiscussionCase-based learning is an effective method and incorporation of it on a regular basis could help favor an effective learning environment.

category: scientific communication and education


title: Functional connectivity of the developing mouse cortex

abstract: Cross-sectional studies have established a variety of structural, synaptic and cell physiological changes corresponding to key critical periods in cortical development. However, the emergence of functional connectivity (FC) in development has not been fully characterized, and hemodynamic-based measures are vulnerable to any neurovascular coupling changes occurring in parallel. We therefore used optical fluorescence imaging to trace longitudinal calcium FC in the awake, resting-state mouse cortex in the same mice at 5 developmental time points beginning at postnatal day 15 (P15) and ending in early adulthood (P60), resulting in over 500 imaging epochs with both calcium and hemodynamics available as a resource for the field. Proof-of-principle analyses revealed that calcium FC displayed coherent functional maps as early as P15, and FC significantly varied in connections between many regions across development, with the developmental trajectorys shape specific to the functional region. This longitudinal developmental calcium FC dataset provides an essential resource for further algorithm development and studies of healthy development and neurodevelopmental disorders.

category: neuroscience


title: Serum CSF1 levels are elevated in patients following partial hepatectomy and acetaminophen induced acute liver failure: Reanalysis of previous data

abstract: A previous study analysed serum CSF1 concentration in patients who had undergone partial hepatectomy and patients with acute liver failure (ALF) induced by acetaminophen. A correlation between low CSF1 and poor prognosis was reported in ALF. In May 2018, an allegation of research misconduct was made against the second author of the published study, D.J.Antoine, who carried out the CSF1 analysis. The current study describes the independent reanalysis of circulating CSF1 concentration in sera from the same patient cohorts. Patient serum CSF1 levels increased following partial hepatectomy. By contrast to the published report, serum CSF1 concentration was greatest in ALF patients who died compared to survivors. The results confirm the relationship between liver injury and circulating serum CSF1.


title: Hybridization and the coexistence of species

abstract: It is thought that two species can coexist if they use different resources present in the environment, yet this assumes that species are completely reproductively isolated. We model coexistence outcomes for two sympatric species that are ecologically differentiated but have incomplete reproductive isolation. The consequences of interbreeding depend crucially on hybrid fitness. When hybrid fitness is high, just a small rate of hybridization can lead to collapse of two species into one. Low hybrid fitness can cause population declines, making extinction of one or both species likely. High intrinsic growth rates result in higher reproductive rates when populations are below carrying capacity, reducing the probability of extinction and increasing the probability of stable coexistence at moderate levels of assortative mating and hybrid fitness. Very strong but incomplete assortative mating can induce low hybrid fitness via a mating disadvantage to rare genotypes, and this can stabilize coexistence of two species at high but incomplete levels of assortative mating. Given these results and evidence that it may take many millions of years of divergence before related species become sympatric, we postulate that coexistence of closely-related species is more often limited by insufficient assortative mating than by insufficient ecological differentiation.


title: Profiling of transcribed cis-regulatory elements in single cells

abstract: Profiling of cis-regulatory elements (CREs, mostly promoters and enhancers) in single cells allows the interrogation of the cell-type and cell-state-specific contexts of gene regulation and genetic predisposition to diseases. Here we demonstrate single-cell RNA-5'end-sequencing (sc-end5-seq) methods can detect transcribed CREs (tCREs), enabling simultaneous quantification of gene expression and enhancer activities in a single assay at no extra cost. We showed enhancer RNAs can be detected using sc-end5-seq methods with either random or oligo(dT) priming. To analyze tCREs in single cells, we developed SCAFE (Single Cell Analysis of Five-prime Ends) to identify genuine tCREs and analyze their activities (https://github.com/chung-lab/scafe). As compared to accessible CRE (aCRE, based on chromatin accessibility), tCREs are more accurate in predicting CRE interactions by co-activity, more sensitive in detecting shifts in alternative promoter usage and more enriched in diseases heritability. Our results highlight additional dimensions within sc-end5-seq data which can be used for interrogating gene regulation and disease heritability.


title: Structure and dynamics of SARS-CoV-2 proofreading exoribonuclease ExoN

abstract: High-fidelity replication of the large RNA genome of coronaviruses (CoVs) is mediated by a 3'-to-5' exoribonuclease (ExoN) in non-structural protein 14 (nsp14), which excises nucleotides including antiviral drugs mis-incorporated by the low-fidelity viral RNA-dependent RNA polymerase (RdRp) and has also been implicated in viral RNA recombination and resistance to innate immunity. Here we determined a 1.6-[A] resolution crystal structure of SARS-CoV-2 ExoN in complex with its essential co-factor, nsp10. The structure shows a highly basic and concave surface flanking the active site, comprising several Lys residues of nsp14 and the N-terminal amino group of nsp10. Modeling suggests that this basic patch binds to the template strand of double-stranded RNA substrates to position the 3' end of the nascent strand in the ExoN active site, which is corroborated by mutational and computational analyses. Molecular dynamics simulations further show remarkable flexibility of multi-domain nsp14 and suggest that nsp10 stabilizes ExoN for substrate RNA-binding to support its exoribonuclease activity. Our high-resolution structure of the SARS-CoV-2 ExoN-nsp10 complex serves as a platform for future development of anti-coronaviral drugs or strategies to attenuate the viral virulence.

category: biochemistry


title: Computational and Image Processing Methods for Analysis and Automation of Anatomical Alignment and Joint Spacing in Reconstructive Surgery

abstract:
PurposeReconstructive surgeries to treat a number of musculoskeletal conditions, from arthritis to severe trauma, involve implant placement and reconstructive planning components. Anatomically matched 3D printed implants are becoming increasingly patient-specific; however the preoperative planning and design process requires several hours of manual effort from highly trained engineers and clinicians. Our work mitigates this problem by proposing algorithms for the automatic re-alignment of unhealthy anatomies, leading to more efficient, affordable, and scalable treatment solutions.

MethodsOur solution combines global alignment techniques such as iterative closest points (ICP) with novel joint space refinement algorithms. The latter is achieved by a low dimensional characterization of the joint space, computed from the distribution of the distance between adjacent points in a joint.

ResultsExperimental validation is presented on real clinical data from human subjects. Compared with ground truth healthy anatomies, our algorithms can reduce misalignment errors by 22% in translation and 19% in rotation for the full foot-and-ankle and 37% in translation and 39% in rotation for the hind-foot only, achieving a performance comparable to expert technicians.

ConclusionOur methods and histogram-based metric allow for automatic and unsupervised alignment of anatomies, a major step toward a fully automated and data driven re-positioning, designing, and diagnosing tool.

category: bioengineering


title: Computer Vision and Deep Learning for Environment-Adaptive Control of Robotic Lower-Limb Exoskeletons

abstract: Robotic exoskeletons require human control and decision making to switch between different locomotion modes, which can be inconvenient and cognitively demanding. To support the development of automated locomotion mode recognition systems (i.e., high-level controllers), we designed an environment recognition system using computer vision and deep learning. We collected over 5.6 million images of indoor and outdoor real-world walking environments using a wearable camera system, of which ~923,000 images were annotated using a 12-class hierarchical labelling architecture (called the ExoNet database). We then trained and tested the EfficientNetB0 convolutional neural network, designed for efficiency using neural architecture search, to predict the different walking environments. Our environment recognition system achieved ~73% image classification accuracy. While these preliminary results benchmark Efficient-NetB0 on the ExoNet database, further research is needed to compare different image classification algorithms to develop an accurate and real-time environment-adaptive locomotion mode recognition system for robotic exoskeleton control.


title: Analysis of variability of functionals of recombinant protein production trajectories based on limited data

abstract: We propose a method for analyzing the variability in smooth, possibly nonlinear, functionals associated with a set of product production trajectories measured under different experimental conditions. The key challenge is to make meaningful inference on these parameters across different experimental conditions when only a limited number of measurements in time are collected for each treatment, and when there are only a few, or no, replicates available. For this purpose we adopt a modeling approach by representing the production trajectories in a B-spline basis, and develop a bootstrap-based inference procedure for the parameters of interest, also accounting for multiple comparisons. The methodology is applied to study two types of quantities of interest - "time to harvest" and "maximal productivity" in the context of an experiment on the production of certain recombinant proteins under laboratory conditions. We complement the findings by extensive numerical experiments that look into the effects of different types of bootstrap procedures and associated schemes for computing p-values for tests of hypotheses.


title: Transport Motility of Phagosomes on Actin and Microtubules Regulates Timing and Kinetics of Their Maturation

abstract: Immune cells degrade internalized pathogens in phagosomes through sequential biochemical changes. The degradation must be fast enough for effective infection control. The presumption is that each phagosome degrades cargos autonomously with a distinct but stochastic kinetic rate. Here we report that the degradation kinetics of individual phagosomes is not stochastic but coupled to their intracellular motility. By engineering RotSensors that are optically anisotropic, magnetic responsive, and fluorogenic in response to degradation activities in phagosomes, we monitored cargo degradation kinetics in single phagosomes simultaneously with their translational and rotational dynamics. We show that phagosomes that move faster centripetally are more likely to encounter and fuse with lysosomes, thereby acidifying faster and degrading cargos more efficiently. The degradation rates increase nearly linearly with the translational and rotational velocities of phagosomes. Our results indicate that the centripetal motion of phagosomes functions as a clock for controlling the progression of cargo degradation.


title: Heterogeneous but not random: Cargo degradation in phagosomes is kinetically coupled to 1 their intracellular mobility


title: THEM6-mediated lipid remodelling sustains stress resistance in cancer

abstract: Despite the clinical benefit of androgen-deprivation therapy (ADT), the majority of patients with advanced prostate cancer (PCa) ultimately develop lethal castration-resistant prostate cancer (CRPC). In this study, we identified thioesterase superfamily member 6 (THEM6) as a marker of ADT resistance in PCa. In patients, THEM6 expression correlates with progressive disease and is associated with poor survival. THEM6 deletion reduces in vivo tumour growth and restores castration sensitivity in orthograft models of CRPC. Mechanistically, THEM6 is located at the endoplasmic reticulum (ER) membrane and controls lipid homeostasis by regulating intracellular levels of ether lipids. Consequently, THEM6 loss in CRPC cells significantly alters ER function, reducing de novo sterol biosynthesis and preventing lipid-mediated induction of ATF4. Finally, we show that THEM6 is required for the establishment of the MYC-induced stress response. Thus, similar to PCa, THEM6 loss significantly impairs tumorigenesis in the MYC-dependent subtype of triple negative breast cancer. Altogether, our results highlight THEM6 as a novel component of the treatment-induced stress response and a promising target for the treatment of CRPC and MYC-driven cancer.


title: Unbiased clade age estimation using a Bayesian Brownian Bridge

abstract: In a recent paper1 we presented a new model, the Bayesian Brownian Bridge (BBB), to infer clade age based on fossil evidence and modern diversity. We benchmarked the method with extensive simulations, including a wide range of diversification histories and sampling heterogeneities that go well beyond the necessarily simplistic model assumptions. Applying BBB to 198 angiosperm families, we found that their fossil record is compatible with clade origins earlier than most contemporary palaeobotanical interpretations. In particular, we estimated with high probability that crown-angiosperms originated before the Cretaceous (> 145 Ma). Budd and colleagues2 critique our study, arguing that the BBB model is biased towards older estimates when fossil data are scarce or absent, that our underlying fossil dataset is unsound, that our clade age estimates are therefore biased by early diverging lineages that are underrepresented in the fossil record, and that pooling of fossil data for analysis at higher taxonomic ranks overcomes these biases. Here, we explore their points and perform new simulations to show that their critique has no merit.

category: paleontology


title: Ozone Stress Responsive Gene Database (OSRGD ver. 1.1): a literature curated database for crop improvement

abstract: Ozone (O3) is a major abiotic stress which severely affects the growth and development of plants. In order to cope up with ozone stress, plants exhibit a plethora of morphological, physiological and molecular changes. Various molecular studies have been performed and a variety of genes exhibiting expression in response to ozone stress have been identified. However, no existing database has been designed yet to present information on ozone stress responsive genes differentially expressed across different plant species. The lack of a curated database limits the research potential in the area and therefore a cohesive database should be designed as a data repository of the ozone responsive genes. OSRGD ver. 1.1 is a user friendly web interface to explore ozone stress specific transcriptome dataset of different plant species. It allows users to retrieve the ozone stress responsive gene data by keyword based search. Each entry upon keyword query contains detailed information about the specific species, its foliar injury symptoms, pattern of gene expression, and platform used for gene expression analysis with reference literature. This comprehensive biocuration will offer researchers a better biological insight into plants response to ozone stress which will focus on development of climate smart crops through crop breeding.

category: plant biology


title: Phosphorylation-dependent control of Arc protein by synaptic plasticity regulator TNIK

abstract: Activity-regulated cytoskeleton-associated protein (Arc) is an immediate-early gene product that support neuroplastic changes important for cognitive function and memory formation. As a protein with homology to the retroviral Gag protein, a particular characteristic of Arc is its capacity to self-assemble into virus-like capsids that can package mRNAs and transfer those transcripts to other cells. Although a lot has been uncovered about the contributions of Arc to neuron biology and behavior, very little is known about how different functions of Arc are coordinately regulated both temporally and spatially in neurons. The answer to this question we hypothesized must involve the occurrence of different protein post-translational modifications acting to confer specificity. In this study, we used mass spectrometry and sequence prediction strategies to map novel Arc phosphorylation sites. Our approach led us to recognize serine 67 (S67) and threonine 278 (T278) as residues that can be modified by TNIK, which is a kinase abundantly expressed in neurons that shares many functional overlaps with Arc and has, along with its interacting proteins such as the NMDA receptor, been implicated as a risk factor for psychiatric disorders. Furthermore, characterization of each residue using site-directed mutagenesis to create S67 and T278 mutant variants revealed that TNIK action at those amino acids can strongly influence Arcs subcellular distribution and self-assembly as capsids. Together, our findings reveal an unsuspected connection between Arc and TNIK. Better understanding of the interplay between these two proteins in neuronal cells could lead to new insights about apparition and progression of psychiatric disorders.


title: Recovery from acute SARS-CoV-2 infection and development of anamnestic immune responses in T cell-depleted rhesus macaques

abstract: Severe COVID-19 has been associated with T cell lymphopenia 1,2, but no causal effect of T cell deficiency on disease severity has been established. To investigate the specific role of T cells in recovery from SARS-CoV-2 infections we studied rhesus macaques that were depleted of either CD4+, CD8+ or both T cell subsets prior to infection. Peak virus loads were similar in all groups, but the resolution of virus in the T cell-depleted animals was slightly delayed compared to controls. The T cell-depleted groups developed virus-neutralizing antibody responses and also class-switched to IgG. When re-infected six weeks later, the T cell-depleted animals showed anamnestic immune responses characterized by rapid induction of high-titer virus-neutralizing antibodies, faster control of virus loads and reduced clinical signs. These results indicate that while T cells play a role in the recovery of rhesus macaques from acute SARS-CoV-2 infections, their depletion does not induce severe disease, and T cells do not account for the natural resistance of rhesus macaques to severe COVID-19. Neither primed CD4+ or CD8+ T cells appeared critical for immunoglobulin class switching, the development of immunological memory or protection from a second infection.

category: immunology


title: Distribution, organization and expression of genes concerned with anaerobic lactate-utilization in human intestinal bacteria

abstract:
Lactate accumulation in the human gut is linked to a range of deleterious health impacts. However, lactate is consumed and converted to the beneficial short chain fatty acids butyrate and propionate by indigenous lactate-utilizing bacteria. To better understand the underlying genetic basis for lactate utilization, transcriptomic analysis was performed for two prominent lactate-utilizing species from the human gut, Anaerobutyricum soehngenii and Coprococcus catus, during growth on lactate, hexose sugar, or hexose plus lactate. In A. soehngenii L2-7, six genes of the lct cluster including NAD-independent D-lactate dehydrogenase (i-LDH) were co-ordinately upregulated during growth on equimolar D and L-lactate (DL-lactate). Upregulated genes included an acyl-CoA dehydrogenase related to butyryl-CoA dehydrogenase, which may play a role in transferring reducing equivalents between reduction of crotonyl-CoA and oxidation of lactate. Genes upregulated in C. catus GD/7 included a six-gene cluster (lap) encoding propionyl CoA-transferase, a putative lactoyl-CoA epimerase, lactoyl-CoA dehydratase and lactate permease, and two unlinked acyl-CoA dehydrogenase genes that are candidates for acryloyl-CoA reductase. An i-LDH homolog in C. catus is encoded by a separate, partial lct, gene cluster, but not upregulated on lactate. While C. catus converts three mols of DL-lactate via the acrylate pathway to two mols propionate and one mol acetate, some of the acetate can be re-used with additional lactate to produce butyrate. A key regulatory difference is that while glucose partially repressed lct cluster expression in A. soehngenii, there was no repression of lactate utilization genes by fructose in the non-glucose utilizer C. catus. This implies that bacteria such as C. catus might be more important in curtailing lactate accumulation in the gut.

Impact statementLactate can be produced as a fermentation by-product by many gut bacteria but has the potential to perturb intestinal microbial communities by lowering luminal pH, and its accumulation has been linked to a range of deleterious health outcomes. Fortunately, in healthy individuals, lactate tends not to accumulate as it is consumed by cross-feeding lactate-utilizing bacteria, which can convert it into the beneficial short chain fatty acids butyrate and propionate. Lactate-utilizing gut bacteria are therefore promising candidates for potential development as novel probiotics. However, lactate-utilizers are taxonomically diverse, and the genes that underpin utilization of lactate by these specialized gut bacteria are not fully understood. In this study we used transcriptomics to compare gene expression profiles of Anaerobutyricum soehngenii and Coprococcus catus, two prominent lactate-utilizing species in the human gut, during growth on lactate alone, sugar alone, or sugar plus lactate. The results revealed strong upregulation of key, but distinct, gene clusters that appear to be responsible for lactate utilization by these, and other, gut bacterial species. Our results therefore increase mechanistic understanding of different lactate utilization pathways used by gut bacteria, which may help to inform selection of optimal lactate-utilizing species for development as novel therapeutics against colonic microbiota perturbations.

Data summaryNovel draft genomes generated for this study have been made available from GenBank (https://www.ncbi.nlm.nih.gov/bioproject/) under BioProject number PRJNA701799. RNA-seq data have been deposited in the ArrayExpress database at EMBL-EBI (www.ebi.ac.uk/arrayexpress) under accession number E-MTAB-10136. Further details of additional existing genomic data that were analyzed in this project are given in Table 1 and Table S2.

O_TBL View this table:
org.highwire.dtl.DTLVardef@13fd07borg.highwire.dtl.DTLVardef@90796dorg.highwire.dtl.DTLVardef@1140a60org.highwire.dtl.DTLVardef@b53597org.highwire.dtl.DTLVardef@d16268_HPS_FORMAT_FIGEXP  M_TBL O_FLOATNOTable 1.C_FLOATNO O_TABLECAPTIONNew and existing strains used for in vitro cultivation studies in this work, and their corresponding genomes. "N/A" indicates no genome available for this strain, which was used for in vitro work only. The full list of strains used for genomic-based analyses is shown in Table S2.

C_TABLECAPTION C_TBL

category: microbiology


title: Membrane lectins enhance SARS-CoV-2 infection and influence the neutralizing activity of different classes of antibodies

abstract: Investigating the mechanisms of SARS-CoV-2 cellular infection is key to better understand COVID-19 immunity and pathogenesis. Infection, which involves both cell attachment and membrane fusion, relies on the ACE2 receptor that is paradoxically found at low levels in the respiratory tract, suggesting that additional mechanisms facilitating infection may exist. Here we show that C-type lectin receptors, DC-SIGN, L-SIGN and the sialic acid-binding Ig-like lectin 1 (SIGLEC1) function as auxiliary receptors by enhancing ACE2-mediated infection and modulating the neutralizing activity of different classes of spike-specific antibodies. Antibodies to the N-terminal domain (NTD) or to the conserved proteoglycan site at the base of the Receptor Binding Domain (RBD), while poorly neutralizing infection of ACE2 over-expressing cells, effectively block lectin-facilitated infection. Conversely, antibodies to the Receptor Binding Motif (RBM), while potently neutralizing infection of ACE2 over-expressing cells, poorly neutralize infection of cells expressing DC-SIGN or L-SIGN and trigger fusogenic rearrangement of the spike promoting cell-to-cell fusion. Collectively, these findings identify a lectin-dependent pathway that enhances ACE2-dependent infection by SARS-CoV-2 and reveal distinct mechanisms of neutralization by different classes of spike-specific antibodies.


title: Sex differences in lung imaging and SARS-CoV-2 antibody responses in a COVID-19 golden Syrian hamster model

abstract:
In the ongoing coronavirus disease 2019 (COVID-19) pandemic caused by the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), more severe outcomes are reported in males compared with females, including hospitalizations and deaths. Animal models can provide an opportunity to mechanistically interrogate causes of sex differences in the pathogenesis of SARS-CoV-2. Adult male and female golden Syrian hamsters (8-10 weeks of age) were inoculated intranasally with 105 TCID50 of SARS-CoV-2/USA-WA1/2020 and euthanized at several time points during the acute (i.e., virus actively replicating) and recovery (i.e., after the infectious virus has been cleared) phases of infection. There was no mortality, but infected male hamsters experienced greater morbidity, losing a greater percentage of body mass, developing more extensive pneumonia as noted on chest computed tomography, and recovering more slowly than females. Treatment of male hamsters with estradiol did not alter pulmonary damage. Virus titers in respiratory tissues, including nasal turbinates, trachea, and lungs, and pulmonary cytokine concentrations, including IFN{beta} and TNF, were comparable between the sexes. However, during the recovery phase of infection, females mounted two-fold greater IgM, IgG, and IgA responses against the receptor-binding domain of the spike protein (S-RBD) in both plasma and respiratory tissues. Female hamsters also had significantly greater IgG antibodies against whole inactivated SARS-CoV-2 and mutant S-RBDs, as well as virus neutralizing antibodies in plasma. The development of an animal model to study COVID-19 sex differences will allow for a greater mechanistic understanding of the SARS-CoV-2 associated sex differences seen in the human population.

ImportanceMen experience more severe outcomes from COVID-19 than women. Golden Syrian hamsters were used to explore sex differences in the pathogenesis of a human clinical isolate of SARS-CoV-2. After inoculation, male hamsters experienced greater sickness, developed more severe lung pathology, and recovered more slowly than females. Sex differences in disease could not be reversed by estradiol treatment in males and were not explained by either virus replication kinetics or the concentrations of inflammatory cytokines in the lungs. During the recovery period, antiviral antibody responses in the respiratory tract and plasma, including to newly emerging SARS-CoV-2 variants, were greater in females than male hamsters. Greater lung pathology during the acute phase combined with reduced antiviral antibody responses during the recovery phase of infection in males than females illustrate the utility of golden Syrian hamsters as a model to explore sex differences in the pathogenesis of SARS-CoV-2 and vaccine-induced immunity and protection.

One Sentence SummaryFollowing SARS-CoV-2 infection, male hamsters experience worse clinical disease and have lower antiviral antibody responses than females.


title: Sterilizing Effects of Novel Regimens Containing TB47, Clofazimine and Linezolid in a Murine Model of Tuberculosis.

abstract: Toxicity and inconvenience associated with the use of injectable drug-containing regimens for tuberculosis (TB) have made all-oral regimens a preferred alternative. Widespread resistance to fluoroquinolones and pyrazinamide makes it essential to identify new drug candidates and study their effects on current regimens for TB. TB47 is a pyrazolo[1,5-a]pyridine-3-carboxamide with powerful synergistic in vitro and in vivo activities against mycobacteria, especially with clofazimine. Here, we investigated the bactericidal and sterilizing activities of novel oral regimens containing TB47 + clofazimine + linezolid, and the potential roles of levofloxacin and/or pyrazinamide in such drug combinations. Using a well-established mouse model, we assessed the effect of these regimens on bacterial burden in the lung during treatment and relapse (4 months after stopping treatment + immunosuppression). Our findings indicate that the TB47 + clofazimine + linezolid + pyrazinamide, with/without levofloxacin, regimens had fast-acting (4 months) sterilizing activity and no relapse was observed. When pyrazinamide was excluded from the regimen, treatment times were longer (5-6 months) to achieve sterilizing conditions. We propose that TB47 + clofazimine + linezolid can form a highly sterilizing block for use as an alternative pan-TB regimen.


title: The most recent common ancestor for Y chromosome lived about 3.67 million years ago

abstract: Investigating the divergence time between different populations has been of fundamental interest in evolution biology. In this study, we reanalysed the mitochondrial and Y chromosome DNA of the 1000 Genomes Project, and revealed that most single nucleotide polymorphisms (SNPs) are specific to minority and would be classified as low-frequency mutations. Using these polymorphisms, we recalculated the divergence time of different populations by direct comparison between individuals and the out-group.

category: evolutionary biology


title: Calculating the divergence time by direct comparison between individuals and the out-group


title: Agronomic behavior of four cultivars and 56 F1 tomato populations

abstract: This research was conducted under a greenhouse at the National Vegetable Seed Production Center located in the municipality of Sipe Sipe, Quillacollo province with the aim of analyzing agronomic behavior in 56 F1 progenies and four hybrid tomato cultivars. The research consisted of 60 genotypes as treatments. Each experimental unit was made up of 6 plants. The variables evaluated were: number of flowers per inflorescence, number of fruits per cluster, weight of fruit, equatorial diameter, polar diameter, heart diameter, thickness of pericarp, number of fruits per plant, weight of fruit per plant, hardness and solids soluble . The data obtained were analyzed with descriptive statistics, correlation, cluster and principal component analysis. Significant correlations were found for the number of fruits per plant and number of flowers per inflorescence with the weight of fruit per plant. The polar dendogram identified four distant groups in the performance and quality components. The principal component analysis identified the degree of association between variables and genotypes, identifying groups correlated to components of fruit yield and quality.

category: genetics


title: Genomic resources for the North American water vole (Microtus richardsoni) and the montane vole (Microtus montanus)

abstract:
BackgroundVoles of the genus Microtus are important research organisms, yet genomic resources in the genus are lacking. Providing such resources would benefit future studies of immunology, phylogeography, cryptic diversity, and more.

FindingsWe sequenced and assembled nuclear genomes from two subspecies of water vole (Microtus richardsoni) and from the montane vole (Microtus montanus). The water vole genomes were sequenced with Illumina and 10X Chromium plus Illumina sequencing, resulting in assemblies with ~1,600,000 and ~30,000 scaffolds respectively. The montane vole was assembled into ~13,000 scaffolds using Illumina sequencing also. In addition to the nuclear assemblies, mitochondrial genome assemblies were also performed for both species. We conducted a structural and functional annotation for the best water vole nuclear genome, which resulted in ~24,500 annotated genes, with 83% of these receiving functional annotations. Finally, we find that assembly quality statistics for our nuclear assemblies fall within the range of genomes previously published in the genus Microtus, making the water vole and montane vole genomes useful additions to currently available genomic resources.

category: genomics


title: A Tensegrity-Graph Optimization Framework for Estimating Intracellular Stiffness

abstract: In this paper we present a formulation for estimating intracellular stiffness using tensegrity principles. We demonstrate that the new tensegrity model, based on random graphs over dense nodes, can predict well known mechanical characteristics of epithelial and mesenchymal cells.


title: Transport of Ketoprofen in Mammalian Blood Plasma

abstract: Ketoprofen is a popular non-steroidal anti-inflammatory drug (NSAID) transported in the bloodstream mainly by serum albumin (SA). Ketoprofen is known to have multiple side effects and interactions with hundreds of other drugs, which might be related to its vascular transport by SA. Our work reveals that ketoprofen binds to a different subset of drug binding sites on human SA than has been observed for other species, despite the conservation of drug sites between species. We discuss potential reasons for the observed differences in the drugs preferences for particular sites, including ketoprofen binding determinants in mammalian SAs and the effect of fatty acids on drug binding. The presented results show that the SA drug sites to which a particular drug binds cannot be easily predicted based only on a complex of SA from another species and the conservation of drug sites between species.


title: Inter-chromosomal linkage disequilibrium and linked fitness cost loci influence the evolution of nontarget site herbicide resistance in an agricultural weed

abstract: The adaptation of weedy plants to herbicide is both a significant problem in agriculture and a model for the study of rapid adaptation under regimes of strong selection. Despite recent advances in our understanding of simple genetic changes that lead to resistance, a significant gap remains in our knowledge of resistance controlled by many loci and the evolutionary factors that influence the maintenance of resistance over time. Here, we perform a multi-level analysis involving whole genome sequencing and assembly, resequencing and gene expression analysis to both uncover putative loci involved in nontarget herbicide resistance and to examine evolutionary forces underlying the maintenance of resistance in natural populations. We found loci involved in herbicide detoxification, stress sensing, and alterations in the shikimate acid pathway to be under selection, and confirmed that detoxification is responsible for glyphosate resistance using a functional assay. Furthermore, we found interchromosomal linkage disequilibrium (ILD), most likely associated with epistatic selection, to influence NTSR loci found on separate chromosomes thus potentially mediating resistance through generations. Additionally, by combining the selection screen, differential expression and LD analysis, we identified fitness cost loci that are strongly linked to resistance alleles, indicating the role of genetic hitchhiking in maintaining the cost. Overall, our work strongly suggests that NTSR glyphosate resistance in I. purpurea is conferred by multiple genes which are maintained through generations via ILD, and that the fitness cost associated with resistance in this species is a by-product of genetic-hitchhiking.


title: Inter-chromosomal linkage disequilibrium and linked fitness cost loci associated with selection for herbicide resistance


title: A New Genus Of Microsporidian Parasite (Hepatosporidae; Microsporidia) Found In The Oocytes Of Ribbon Worms From The North Pacific Genus Maculaura (Heteronemertea; Nemertea)

abstract:
An intracellular microsporidian parasite was first observed within oocytes of Maculaura alaskensis, a small pilidiophoran nemertean, commonly found on sandflats along the Pacific coast of North America. Infected oocytes have large vesicles containing dozens to hundreds of diplokaryotic, ellipsoid spores measuring 1.3 by 2.3 m. A partial small subunit nuclear ribosomal 18S gene sequence isolated from the microsporidian does not match any known microsporidian sequences in the public databases. Phylogenetic analysis groups it with Hepatospora eriocheir in a sister clade to the Enterocytozoonidae. All the known life stages of this parasite are contained within a membranous envelope. This microsporidian was identified in M. alaskensis, Maculaura aquilonia, Maculaura oregonensis, and Maculaura cerebrosa in Coos Bay, Oregon, in M. alaskensis from Newport, Oregon, and in M. aquilonia collected in Juneau, Alaska. This is, to our knowledge, the first species of microsporidia found to directly infect nemertean host cells.

Graphical abstract

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category: zoology


title: Deep Generative Analysis for Task-Based Functional MRI Experiments

abstract: While functional magnetic resonance imaging (fMRI) remains one of the most widespread and important methods in basic and clinical neuroscience, the data it produces--time series of brain volumes--continue to pose daunting analysis challenges. The current standard ("mass univariate") approach involves constructing a matrix of task regressors, fitting a separate general linear model at each volume pixel ("voxel"), computing test statistics for each model, and correcting for false positives post hoc using bootstrap or other resampling methods. Despite its simplicity, this approach has enjoyed great success over the last two decades due to: 1) its ability to produce effect maps highlighting brain regions whose activity significantly correlates with a given variable of interest; and 2) its modeling of experimental effects as separable and thus easily interpretable. However, this approach suffers from several well-known drawbacks, namely: inaccurate assumptions of linearity and noise Gaussianity; a limited ability to capture individual effects and variability; and difficulties in performing proper statistical testing secondary to independently fitting voxels. In this work, we adopt a different approach, modeling entire volumes directly in a manner that increases model flexibility while preserving interpretability. Specifically, we use a generalized additive model (GAM) in which the effects of each regressor remain separable, the product of a spatial map produced by a variational autoencoder and a (potentially nonlinear) gain modeled by a covariate-specific Gaussian Process. The result is a model that yields group-level effect maps comparable or superior to the ones obtained with standard fMRI analysis software while also producing single-subject effect maps capturing individual differences. This suggests that generative models with a decomposable structure might offer a more flexible alternative for the analysis of task-based fMRI data.


title: Mixed selectivity coding of sensory and motor social signals in the thalamus of a weakly electric fish

abstract: Recent studies have shown that high-level neural activity often exhibits mixed selectivity to multivariate signals. How such representations arise and how they modulate natural behavior is poorly understood. The social behavior of weakly electric fish is relatively low-dimensional and easily reproduced in the laboratory. Here we show how electrosensory signals related to courtship and rivalry in Apteronotus leptorhynchus are represented in the preglomerular nucleus, the thalamic region exclusively connecting the midbrain with the pallium. We show that preglomerular cells convert their midbrain inputs into a mixed selectivity code that includes corollary discharge of outgoing communication signals. We discuss how the preglomerular pallial targets might use these inputs to control social behavior and determine dominance in male-male competition and female mate selection during courtship. Our results showcase the potential of the electrocommunication system as an accessible model for studying the neural substrates of social behavior and principles of multi-dimensional neural representation.


title: PRMT5 regulates ovarian follicle development by facilitating Wt1 translation

abstract: Protein arginine methyltransferase 5 (Prmt5) is the major type II enzyme responsible for symmetric dimethylation of arginine. Here, we found PRMT5 was expressed at high level in ovarian granulosa cells of growing follicles. Inactivation of Prmt5 in granulosa cells resulted in aberrant follicle development and female infertility. In Prmt5-knockout mice, follicle development was arrested with disorganized granulosa cells in which WT1 expression was dramatically reduced and the expression of steroidogenesis-related genes was significantly increased. The premature differentiated granulosa cells were detached from oocytes and follicle structure was disrupted. Mechanism studies revealed that Wt1 expression was regulated by PRMT5 at the protein level. PRMT5 facilitated IRES-dependent translation of Wt1 mRNA by methylating HnRNPA1. Moreover, the upregulation of steroidogenic genes in Prmt5-deficient granulosa cells was repressed by Wt1 overexpression. These results demonstrate PRMT5 participates in granulosa cell lineage maintenance by inducing Wt1 expression. Our study uncovers a new role of post-translational arginine methylation in granulosa cell differentiation and follicle development.


title: Transcriptomes across fertilization and seed development in the water lily Nymphaea thermarum (Nymphaeales) reveal dynamic expression of DNA and histone methylation modifiers

abstract: Studies of gene expression during seed development have been performed for a growing collection of species from a phylogenetically broad sampling of flowering plants (angiosperms). However, attention has mostly been focused on crop species or a small number of  model systems. Information on gene expression during seed development is minimal for those angiosperm lineages whose origins predate the divergence of monocots and eudicots. In order to provide a new perspective on the early evolution of seed development in flowering plants, we sequenced transcriptomes of whole ovules and seeds from three key stages of reproductive development in the waterlily Nymphaea thermarum, an experimentally-tractable member of the Nymphaeales. We first explore general patterns of gene expression, beginning with mature ovules and continuing through fertilization into early- and mid-seed development. We then examine the expression of genes associated with DNA and histone methylation - processes known to be essential for development in distantly-related and structurally-divergent monocots and eudicots. Around 60% of transcripts putatively homologous to DNA and histone methylation modifiers are differentially expressed during seed development in N. thermarum, suggesting that the importance of dynamic epigenetic patterning during seed development dates to the earliest phases of angiosperm evolution. However, genes involved in establishing, maintaining, and removing methylation marks associated with genetic imprinting show a mix of conserved and unique expression patterns between N. thermarum and other flowering plants. Our data suggests that the regulation of imprinting has likely changed throughout angiosperm evolution, and furthermore identifies genes that merit further characterization in any angiosperm system.


title: Differential neural activity patterns mediate learning across contexts in a social cichlid fish

abstract: Learning and decision-making are greatly influenced by the social context surrounding individuals. When navigating a complex social world, individuals must quickly ascertain where to gain important resources and which group members are useful sources of such information. Such dynamic behavioral processes require neural mechanisms that are flexible across contexts. Here we examined how the social context influences the learning response during a visual cue discrimination task and the neural activity patterns that underlie acquisition of this novel information. Using the cichlid fish, Astatotilapia burtoni, we show that learning of the task is faster in social groups than in a non-social context. We quantified the expression of Fos, an immediate-early gene, across candidate brain regions known to play a role in social behavior and learning, such as the putative teleost homologues of the mammalian hippocampus, basolateral amygdala, and medial amygdala/BNST complex. We found that neural activity patterns differ between social and non-social contexts. Our results suggest that while the same brain regions may be involved in the learning of a discrimination task independent of social context, activity in each region encodes specific aspects of the task based on context.


title: What is the role of the environment in the emergence of novel antibiotic resistance genes? - A modelling approach

abstract: It is generally accepted that intervention strategies to curb antibiotic resistance cannot solely focus on human and veterinary medicine but must also consider environmental settings. While the environment clearly has a role in the transmission of resistant bacteria, it is less clear what role it plays in the emergence of novel types of resistance. It has been suggested that the environment constitutes an enormous recruitment ground for resistance genes to pathogens, but the extent to which this actually happens is unknown. In this study, we built a model framework for resistance emergence and used the available quantitative data on the relevant processes to identify the steps which are limiting the appearance of antibiotic resistance determinants in human or animal pathogens. We also assessed the effect of uncertainty in the available data on the model results. We found that in a majority of scenarios, the environment would only play a minor role in the emergence of novel resistance genes. However, the uncertainty around this role is enormous, highlighting an urgent need of more quantitative data to understand the role of the environment in antibiotic resistance development. Specifically, more data is most needed on the fitness costs of antibiotic resistance gene (ARG) carriage, the degree of dispersal of resistant bacteria from the environment to humans, but also the rates of mobilization and horizontal transfer of ARGs. Quantitative data on these processes is instrumental to determine which processes that should be targeted for interventions to curb development and transmission of resistance.


title: Collective border cell migration requires the zinc transporter Catsup to limit endoplasmic reticulum stress

abstract: Collective cell migration is critical for normal development, wound healing, and in tumor progression and metastasis. Border cells in the Drosophila ovary provide a genetically tractable model to identify molecular mechanisms that drive this important cell behavior. In an unbiased screen for defects in border cell migration in mosaic clones, we identified a mutation in the catsup gene. Catsup, the Drosophila ortholog of Zip7, is a large, multifunctional, transmembrane protein of the endoplasmic reticulum (ER), which has been reported to negatively regulate catecholamine biosynthesis, to regulate Notch signaling, to function as a zinc transporter, and to limit ER stress. Here we report that catsup knockdown caused ER stress in border cells and that ectopic induction of ER stress was sufficient to block migration. Notch and EGFR trafficking were also disrupted. Wild type Catsup rescued the migration defect but point mutations known to disrupt the zinc ion transport of Zip7 did not. We conclude that migrating cells are particularly susceptible to defects in zinc transport and ER homeostasis.


title: Bioinformatics analysis of SARS-CoV-2 RBD mutant variants and insights into antibody and ACE2 receptor binding

abstract: Prevailing COVID-19 vaccines are based on the spike protein of earlier SARS-CoV-2 strain that emerged in Wuhan, China. The continuously evolving nature of SARS-CoV-2 resulting emergence of new variants raises the risk of immune absconds. During the last few months, several RBD (receptor-binding domain) variants have been reported to affect the vaccine efficacy considerably. Soon after reporting of a new double mutant variant (L452R & E484Q) in India, the country facing a deadlier second wave of infections which prompts researchers to suspects this variant to be accountable. To address the relevant concerns about this new variant affecting vaccine efficacy, we performed molecular simulation dynamics based structural analysis of spike protein of double mutant (L452R & E484Q) along with K417G variants and earlier reported RBD variants and found structural changes in RBD region after comparing with the wild type. Comparison of the binding affinity of the double mutant and earlier reported RBD variant for ACE2 (angiotensin 2 altered enzymes) receptor and CR3022 antibody with the wild-type strain revealed the lowest binding affinity of the double mutant for CR3022 among all other variants. These findings suggest that the newly emerged double mutant could significantly reduce the impact of the current vaccine which threatens the protective efficacy of current vaccine therapy.

category: bioinformatics


title: The accuracy of NMR protein structures in the Protein Data Bank

abstract: We recently described a method, ANSURR, for measuring the accuracy of NMR protein structures. It is based on comparing residue-specific measures of rigidity from backbone chemical shifts via the random coil index, and from structures. Here, we report the use of ANSURR to analyse NMR ensembles within the Protein Data Bank (PDB). NMR structures cover a wide range of accuracy, which improved over time until about 2005, since when accuracy has not improved. Most structures have accurate secondary structure, but are too floppy, particularly in loops. There is a need for more experimental restraints in loops. The best current accuracy measures are Ramachandran distribution and number of NOE restraints per residue. The precision of structure ensembles correlates with accuracy, as does the number of hydrogen bond restraints per residue. If a structure contains additional components (such as additional polypeptide chains or ligands), then their inclusion improves accuracy. Analysis of over 7000 PDB NMR ensembles is available via our website ansurr.com.

category: biophysics


title: MICOS and F1FO-ATP synthase crosstalk is a fundamental property of mitochondrial cristae

abstract:
Mitochondrial cristae are polymorphic invaginations of the inner membrane that are the fabric of cellular respiration. Both the Mitochondrial Contact Site and Cristae Organization System (MICOS) and the FOF1-ATP synthase are vital for sculpting cristae by opposing membrane bending forces. While MICOS promotes negative curvature at cristae junctions, dimeric FOF1- ATP synthase is crucial for positive curvature at cristae rims. Crosstalk between these two complexes has been observed in bakers yeast, the model organism of the Opisthokonta supergroup. Here, we report that this property is conserved in Trypanosoma brucei, a member of the Discoba supergroup that separated from Opisthokonta [~]2 billion years ago. Specifically, one of the paralogs of the core MICOS subunit Mic10 interacts with dimeric F1FO-ATP synthase, whereas the other core Mic60 subunit has a counteractive effect on F1FO- ATP synthase oligomerization. This is evocative of the nature of MICOS-F1FO-ATP synthase crosstalk in yeast, which is remarkable given the diversification these two complexes have undergone during almost 2 eons of independent evolution. Furthermore, we identified a highly diverged trypanosome homolog of subunit e, which is essential for the stability of F1FO-ATP synthase dimers in yeast. Just like subunit e, it is preferentially associated with dimers, interacts with Mic10 and its silencing results in severe defects to cristae and disintegration of F1FO-ATP synthase dimers. Our findings indicate that crosstalk between MICOS and dimeric F1FO-ATP synthase is a fundamental property impacting cristae shape throughout eukaryotes.

ImportanceMitochondria have undergone profound diversification in separate lineages that have radiated since the last common ancestor of eukaryotes some eons ago. Most eukaryotes are unicellular protists, including etiological agents of infectious diseases like Trypanosoma brucei. Thus, the study of a broad range of protists can reveal fundamental features shared by all eukaryotes and lineage-specific innovations. Here we report that two different protein complexes, MICOS and F1FO-ATP synthase, known to affect mitochondrial architecture, undergo crosstalk in T. brucei, just as in bakers yeast. This is remarkable considering that these complexes have otherwise undergone many changes during their almost two billion years of independent evolution. Thus, this crosstalk is a fundamental property needed to maintain proper mitochondrial structure even if the constituent players considerably diverged.

category: cell biology


title: Single-molecule imaging of cytoplasmic dynein reveals the mechanism of motor activation and cargo capture

abstract: Cytoplasmic dynein 1 (dynein) is the primary minus end-directed motor protein in most eukaryotic cells. Dynein remains in an inactive conformation until the formation of a tripartite complex comprising dynein, its regulator dynactin and a cargo adaptor. How this process of dynein activation occurs is un-clear, since it entails the formation of a three-protein complex inside the crowded environs of a cell. Here, we employed live-cell, single-molecule imaging to visualise and track fluorescently tagged dynein. First, we observed that only [~]30% of dynein molecules that bound to the microtubule (MT) engaged in minus end-directed movement, and that too for a short duration of [~]0.6 s. Next, using high-resolution imaging in live and fixed cells, and using correlative light and electron microscopy, we discovered that dynactin remained persistently attached to MTs, and endosomal cargo remained in proximity to the MTs and dynactin. Finally, we employed two-colour imaging to visualise cargo movement effected by single motor binding. We then used these discoveries as the basis for a stochastic model incorporating dynamic motors binding to cargo located along MTs, and also developed a coarse-grained 3-state run-and-tumble particle (RTP) model for the cargo that quantitatively recapitulates the emergent statistics of cargo movement. Taken together, we discovered a search mechanism that is facilitated by dyneins frequent MT binding-unbinding kinetics: (1) in a futile event when dynein does not encounter cargo anchored in proximity to the MT, dynein dissociates and diffuses into the cytoplasm, (2) when dynein encounters cargo and dynactin upon MT-binding, it moves cargo in a short run. Several of these short runs are undertaken in succession for long-range directed movement. In conclusion, we demonstrate that dynein activation and cargo capture are coupled in a step that relies on the reduction of dimensionality to enable minus end-directed transport in cellulo, and that complex cargo behaviour emerges from stochastic motor-cargo interactions.


title: Single-molecule imaging of cytoplasmic dynein in cellulo reveals the mechanism of motor activation and cargo capture


title: Functional amyloids in the microbiomes of a rat Parkinson's disease model and wild-type rats

abstract:
Cross-seeding between amyloidogenic proteins in the gut is receiving increasing attention as a possible mechanism for initiation or acceleration of amyloid formation by aggregation-prone proteins such as SN, which is central in the development of Parkinsons disease. This is particularly pertinent in view of the growing number of functional (i.e. benign and useful) amyloid proteins discovered in bacteria. Here we identify two functional amyloid proteins, Pr12 and Pr17, in fecal matter from Parkinsons disease transgenic rats and their wild type counterparts, based on their stability against dissolution by formic acid. Both proteins show robust aggregation into ThT-positive aggregates that contain higher-order {beta}-sheets and have a fibrillar morphology, indicative of amyloid proteins. In addition, Pr17 aggregates formed in vitro showed significant resistance against formic acid, suggesting an ability to form highly stable amyloid. Treatment with proteinase K revealed a protected core of approx. 9 kDa. Neither Pr12 nor Pr17, however, affected SN aggregation in vitro. Thus, amyloidogenicity does not per se lead to an ability to cross-seed fibrillation of SN. Our results support the use of proteomics and formic acid to identify amyloid protein in complex mixtures and indicates the existence of numerous functional amyloid proteins in microbiomes.

IMPORTANCEThe bacterial microbiome in the gastrointestinal tract is increasingly seen as important for human health and disease. One area of particular interest is that of neurodegenerative diseases such as Parkinsons which involve pathological aggregation into amyloid of human proteins such as - synuclein (SN). Bacteria are known to form benign or functional amyloid, some of which may initiate unwanted aggregation of e.g. SN in the enteric nervous system through cross-seeding via contact with the microbiome. Here we show that the rat microbiome contains several proteins which form this type of amyloid aggregate both in vivo and in vitro. Although the two proteins we investigate in depth do not directly promote SN aggregation, our work shows that the microbiome potentially harbors a significant number of bacterial amyloid which could play a role in human physiology at various levels.


title: Altered metal distribution in the sr45-1 Arabidopsis mutant causes developmental defects

abstract:
The plant SR (serine/arginine-rich) splicing factor SR45 plays important roles in several biological processes, such as splicing, DNA methylation, innate immunity, glucose regulation and ABA signaling. A homozygous Arabidopsis sr45-1 null mutant is viable, but exhibits diverse phenotypic alterations, including delayed root development, late flowering, shorter siliques with fewer seeds, narrower leaves and petals, and unusual numbers of floral organs. Here, we report that the sr45-1 mutant presents an unexpected constitutive iron deficiency phenotype characterized by altered metal distribution in the plant. RNA-Sequencing highlighted severe perturbations in metal homeostasis, phenylpropanoid pathway, oxidative stress responses, and reproductive development. Ionomic quantification and histochemical staining revealed strong iron accumulation in the sr45-1 root tissues accompanied by an iron starvation in aerial parts. We showed that some sr45-1 developmental abnormalities can be complemented by exogenous iron supply. Our findings provide new insight into the molecular mechanisms governing the phenotypes of the sr45-1 mutant.

One sentence summaryThe sr45-1 mutation affects Fe homeostasis, which results in reproductive defects


title: High performance microbial opsins for spatially and temporally precise perturbations of large neuronal networks

abstract: Patterned optogenetic activation of defined neuronal populations in the intact brain can reveal fundamental aspects of the neural codes of perception and behavior. The biophysical properties of existing optogenetic tools, however, constrain the scale, speed, and fidelity of precise optical control. Here we use structure-guided mutagenesis to engineer opsins that exhibit very high potency while retaining fast kinetics. These new opsins enable large-scale, temporally and spatially precise control of population neural activity in vivo and in vitro. We benchmark these new opsins against existing optogenetics tools with whole-cell electrophysiology and all-optical physiology and provide a detailed biophysical characterization of a diverse family of microbial opsins under two-photon illumination. This establishes a toolkit and a resource for matching the optimal opsin to the goals and constraints of patterned optogenetics experiments. Finally, by combining these new opsins with optimized procedures for cell-specific holographic photo-stimulation, we demonstrate the simultaneous co-activation of several hundred spatially defined neurons with a single hologram, and nearly double that number by temporally interleaving holograms at fast rates. These newly engineered opsins substantially extend the capabilities of patterned illumination optogenetic paradigms for addressing neural circuits and behavior.


title: EgGLUT1 is crucial for the viability of larvae of Echinococcus granulosus sensus lato by involving its glucose uptake

abstract: Cystic echinococcosis (CE) is a zoonotic parasitic disease caused by infection with the larvae of Echinococcus granulosus sensu lato (s.l.) cluster. It is urgent to identify novel drug targets and develop new drug candidates against CE. Glucose transporter 1 (GLUT1) is mainly responsible for the transmembrane transport of glucose to maintain its constant cellular availability and is a recent research hotspot as a drug target in various diseases. However, presence and role of GLUT1 in E. granulosus s.l. (EgGLTU1) was unknown. In this study, we cloned a conserved GLUT1 homology gene (named EgGLUT1-ss) from E. granulosus sensu stricto (s.s.) and found EgGLUT1-ss was crucial for glucose uptake of the protoscoleces of E. granulosus s.s.. WZB117, a GLUT1 inhibitor, inhibited glucose uptake of E. granulosus s.s. and the viability of the metacestode in vitro. In addition, WZB117 showed potent therapeutic activity in E. granulosus s.s.-infected mice: a 10 mg/kg dose of WZB117 significantly reduced the number and weight of parasite cysts as well as the reference drug, albendazole. Our data have defined EgGLUT1 as a key E. granulosus s.l. vulnerability target, involved in its glucose uptake from the host; this opens a new avenue to identify drugs with an ideal activity profile for the treatment of CE.


title: Heterogeneity of Staphylococcus epidermidis in prosthetic joint infections: Time to reevaluate microbiological criteria?

abstract:
Prosthetic joint infection (PJI) is a feared complication after arthroplasty with Staphylococcus epidermidis as a major pathogen. One diagnostic criteria for PJI diagnosis is the finding of phenotypically identical organisms based on common laboratory tests in two or more periprosthetic microbial cultures. Because of phenotypical variation within a genetic clone, and clonal variation within a phenotype, the criteria may be ambiguous. Here, we investigate the extent of diversity among coagulase-negative staphylococci in PJI and characterize in detail S. epidermidis isolates from these infections.

We performed a retrospective cohort study of 62 consecutive patients with PJI caused by coagulase-negative staphylococci (CoNS) from two hospitals in Northern Sweden. From 16 of the patients, two to nine S. epidermidis isolates were available for whole-genome analyses. Hospital-adapted multidrug-resistant genetic clones of S. epidermidis were identified in 40/62 (65%) of the PJIs using a combination of analysis by pulsed-field gel electrophoresis and multiple-locus sequence typing. Whole genome sequencing showed presence of multiple sequence types (STs) in seven (7/16, 44%) PJIs. Among isolates of the same ST, within-patient phenotypical variation in antibiotic susceptibility and/or whole-genome antibiotic resistance gene content was frequent (11/16, 69%).

These results highlight the ambiguity of using phenotypical characterization of S. epidermidis as diagnostic criteria in PJI. The results call for larger systematic studies to determine the frequency of CoNS diversity in PJIs, the implications of such diversity for microbiological diagnostics, and for the therapeutic outcome in patients.


title: Physiological function of Flo11p domains and the particular role of amyloid core sequences of this adhesin in Saccharomyces cerevisiae

abstract: Flocculins are a family of glycosylated proteins that provide yeast cells with several properties such as biofilm formation, flocculation, invasive growth or formation of velum. These proteins are similarly organised with a N-terminal (adhesion) domain, a stalk-like central B-domain with several repeats and a C-terminal sequence carrying a cell wall anchor site. They also contain amyloid {beta}-aggregation-prone sequences whose functional role is still unclear. In this work, we show that Flo11p differs from other flocculins by the presence of unique amyloid-forming sequences, whose the number is critical in the formation of adhesion nanodomains under a physical shear force. Using a genome editing approach to identify the function of domains in Flo11p phenotypes, we show that the formation of cellular aggregates whose density increases with the number of amyloid sequences cannot be attributed to a specific domain of Flo11p. The same is true for plastic adhesion and surface hydrophobicity the intensity of which depends mainly on the abundance of Flo11p on the cell surface. In contrast, the N and C domains of Flo11p are essential for invasive growth in agar, whereas a reduction in the number of repeats of the B domain weakens this phenotype. However, expression of FLO11 alone is not sufficient to trigger this invasion phenotype. Finally, we show that this flocculin contributes to the integrity of the cell wall.