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Targeting host factors exploited by multiple viruses could offer broad-spectrum solutions for pandemic preparedness. Seventeen candidates targeting diverse functions emerged in a screen of 4,413 compounds for SARS-CoV-2 inhibitors. We demonstrated that lapatinib and other approved inhibitors of the ErbB family of receptor tyrosine kinases suppress replication of SARS-CoV-2, Venezuelan equine encephalitis virus (VEEV), and other emerging viruses with a high barrier to resistance. Lapatinib suppressed SARS-CoV-2 entry and later stages of the viral life cycle and showed synergistic effect with the direct-acting antiviral nirmatrelvir. We discovered that ErbB1, ErbB2, and ErbB4 bind SARS-CoV-2 S1 protein and regulate viral and ACE2 internalization, and they are required for VEEV infection. In human lung organoids, lapatinib protected from SARS-CoV-2-induced activation of ErbB-regulated pathways implicated in non-infectious lung injury, proinflammatory cytokine production, and epithelial barrier injury. Lapatinib suppressed VEEV replication, cytokine production, and disruption of blood-brain barrier integrity in microfluidics-based human neurovascular units, and reduced mortality in a lethal infection murine model. We validated lapatinib-mediated inhibition of ErbB activity as an important mechanism of antiviral action. These findings reveal regulation of viral replication, inflammation, and tissue injury via ErbBs and establish a proof of principle for a repurposed, ErbB-targeted approach to combat emerging viruses.

High-throughput sequencing (HTS), more specifically RNA sequencing of plant tissues, has become an indispensable tool for plant virologists to detect and identify plant viruses. During the data analysis step, plant virologists typically compare the obtained sequences to reference virus databases. In this way, they are neglecting sequences without homologies to viruses, which usually represent the majority of sequencing reads. We hypothesized that traces of other pathogens might be detected in this unused sequence data. In the present study, our goal was to investigate whether total RNA-seq data, as generated for plant virus detection, is also suitable for the detection of other plant pathogens and pests. As proof of concept, we first analyzed RNA-seq datasets of plant materials with confirmed infections by cellular pathogens in order to check whether these non-viral pathogens could be easily detected in the data. Next, we set up a community effort to re-analyze existing Illumina RNA-seq datasets used for virus detection to check for the potential presence of non-viral pathogens or pests. In total, 101 datasets from 15 participants derived from 51 different plant species were re-analyzed, of which 37 were selected for subsequent in-depth analyses. In 29 of the 37 selected samples (78%), we found convincing traces of non-viral plant pathogens or pests. The organisms most frequently detected in this way were fungi (15/37 datasets), followed by insects (13/37) and mites (9/37). The presence of some of the detected pathogens was confirmed by independent (q)PCRs analyses. After communicating the results, 6 out of the 15 participants indicated that they were unaware of the possible presence of these pathogens in their sample(s). All participants indicated that they would broaden the scope of their bioinformatic analyses in future studies and thus check for the presence of non-viral pathogens. In conclusion, we show that it is possible to detect non-viral pathogens or pests from total RNA-seq datasets, in this case primarily fungi, insects, and mites. With this study, we hope to raise awareness among plant virologists that their data might be useful for fellow plant pathologists in other disciplines (mycology, entomology, bacteriology) as well.

This article discusses the challenges and opportunities identified in the implementation of the Citylab project in Latin America during the period of 2015-2018. The project was funded by the Erasmus+ Key action 2 programme of the European Union. The project aims to innovate teaching for sustainability in higher education institutions through Problem Based Learning (PBL). Opposed to traditional teaching methods, the pedagogical approach of PBL is a learner-centred approach that takes a complex problem as point of departure instead of existing established knowledge. Since application of such learning methods is limited in Latin America, the Citylab project attempts to introduce PBL in the existing curricula of 12 Latin American universities through the implementation and development of interdisciplinary Citylab modules focusing on sustainable urban development. Citylab project will be presented. Third, we highlight some critical issues and success factors experienced during the project. The findings of this paper are based on (1) self-reported questionnaires from the partners at the end of 2017; (2) onsite visits by the authors and expert visits; (3) focus groups, interviews and conversations with project leaders of the participating institutions during the project. Depending on the institution, the project results were varying in terms of innovation and upscaling potential. Critical factors were related to the role of the project leader in the organization, the flexibility of the implementation and cultural differences. Internal regulations created both incentives and disincentives for participation. Competitive elements in the project and available resources for equipment can act as stimulators in some cases. The challenge lies moreover in detecting windows of opportunities for change in order to accomplish curriculum reform and by doing so, pursue continuation of the PBL approach after the project's horizon.

ObjectiveParoxysmal epileptiform abnormalities on electroencephalography (EEG) are the hallmark of epilepsies, but it is uncertain to what extent epilepsy and background EEG oscillations share neurobiological underpinnings. Here, we aimed to assess the genetic correlation between epilepsy and background EEG oscillations.MethodsConfounding factors, including the heterogeneous etiology of epilepsies and medication effects, hamper studies on background brain activity in people with epilepsy. To overcome this limitation, we compared genetic data from a genome-wide association study (GWAS) on epilepsy (n = 12 803 people with epilepsy and 24 218 controls) with that from a GWAS on background EEG (n = 8425 subjects without epilepsy), in which background EEG oscillation power was quantified in four different frequency bands: alpha, beta, delta, and theta. We replicated our findings in an independent epilepsy replication dataset (n = 4851 people with epilepsy and 20 428 controls). To assess the genetic overlap between these phenotypes, we performed genetic correlation analyses using linkage disequilibrium score regression, polygenic risk scores, and Mendelian randomization analyses.ResultsOur analyses show strong genetic correlations of genetic generalized epilepsy (GGE) with background EEG oscillations, primarily in the beta frequency band. Furthermore, we show that subjects with higher beta and theta polygenic risk scores have a significantly higher risk of having generalized epilepsy. Mendelian randomization analyses suggest a causal effect of GGE genetic liability on beta oscillations.SignificanceOur results point to shared biological mechanisms underlying background EEG oscillations and the susceptibility for GGE, opening avenues to investigate the clinical utility of background EEG oscillations in the diagnostic workup of epilepsy.

Human 4-hydroxyphenylpyruvate dioxygenase-like (HPDL) is a putative iron-containing non-heme oxygenase of unknown specificity and biological significance. We report 25 families containing 34 individuals with neurological disease associated with biallelic HPDL variants. Phenotypes ranged from juvenile-onset pure hereditary spastic paraplegia to infantile-onset spasticity and global developmental delays, sometimes complicated by episodes of neurological and respiratory decompensation. Variants included bona fide pathogenic truncating changes, although most were missense substitutions. Functionality of variants could not be determined directly as the enzymatic specificity of HPDL is unknown; however, when HPDL missense substitutions were introduced into 4-hydroxyphenylpyruvate dioxygenase (HPPD, an HPDL orthologue), they impaired the ability of HPPD to convert 4-hydroxyphenylpyruvate into homogentisate. Moreover, three additional sets of experiments provided evidence for a role of HPDL in the nervous system and further supported its link to neurological disease: (i) HPDL was expressed in the nervous system and expression increased during neural differentiation; (ii) knockdown of zebrafish hpdl led to abnormal motor behaviour, replicating aspects of the human disease; and (iii) HPDL localized to mitochondria, consistent with mitochondrial disease that is often associated with neurological manifestations. Our findings suggest that biallelic HPDL variants cause a syndrome varying from juvenile-onset pure hereditary spastic paraplegia to infantile-onset spastic tetraplegia associated with global developmental delays.

Sequencing-based studies have identified novel risk genes associated with severe epilepsies and revealed an excess of rare deleterious variation in less-severe forms of epilepsy. To identify the shared and distinct ultra-rare genetic risk factors for different types of epilepsies, we performed a whole-exome sequencing (WES) analysis of 9,170 epilepsy-affected individuals and 8,436 controls of European ancestry. We focused on three phenotypic groups: severe developmental and epileptic encephalopathies (DEEs), genetic generalized epilepsy (GGE), and non-acquired focal epilepsy (NAFE). We observed that compared to controls, individuals with any type of epilepsy carried an excess of ultra-rare, deleterious variants in constrained genes and in genes previously associated with epilepsy; we saw the strongest enrichment in individuals with DEEs and the least strong in individuals with NAFE. Moreover, we found that inhibitory GABAA receptor genes were enriched for missense variants across all three classes of epilepsy, whereas no enrichment was seen in excitatory receptor genes. The larger gene groups for the GABAergic pathway or cation channels also showed a significant mutational burden in DEEs and GGE. Although no single gene surpassed exome-wide significance among individuals with GGE or NAFE, highly constrained genes and genes encoding ion channels were among the lead associations; such genes included CACNA1G, EEF1A2, and GABRG2 for GGE and LGI1, TRIM3, and GABRG2 for NAFE. Our study, the largest epilepsy WES study to date, confirms a convergence in the genetics of severe and less-severe epilepsies associated with ultra-rare coding variation, and it highlights a ubiquitous role for GABAergic inhibition in epilepsy etiology.

Maize lethal necrosis (MLN) is a maize disease caused by the maize chlorotic mottle virus (MCMV), a potyvirus which causes yield losses of 30–100%. The present study aimed to isolate nanobodies against the MCMV coat protein (CP) for the diagnosis of MLN. MCMV CP expressed in Escherichia coli was used for llama immunization. VHH (i.e. variable heavy domain of heavy chain) gene fragments were prepared from blood drawn from the immunized llama and used to generate a library in E. coli TG1 cells. MCMV specific nanobodies were selected by three rounds of phage display and panning against MCMV CP. The selected nanobodies were finally expressed in E. coli WK6 cells and purified. Eleven MCMV‐specific nanobodies were identified and shown to detect MCMV in infected maize plants. Thus, our results show that nanobodies isolated from llama immunized with MCMV CP can distinguish infected and healthy maize plants, potentially enabling development of affordable MCMV detection protocols. [ABSTRACT FROM AUTHOR]

Emerging evidence indicates that epileptic encephalopathies are genetically highly heterogeneous, underscoring the need for large cohorts of well-characterized individuals to further define the genetic landscape. Through a collaboration between two consortia (EuroEPINOMICS and Epi4K/EPGP), we analyzed exome-sequencing data of 356 trios with the "classical" epileptic encephalopathies, infantile spasms and Lennox Gastaut syndrome, including 264 trios previously analyzed by the Epi4K/EPGP consortium. In this expanded cohort, we find 429 de novo mutations, including de novo mutations in DNM1 in five individuals and de novo mutations in GABBR2, FASN, and RYR3 in two individuals each. Unlike previous studies, this cohort is sufficiently large to show a significant excess of de novo mutations in epileptic encephalopathy probands compared to the general population using a likelihood analysis (p = 8.2 × 10(-4)), supporting a prominent role for de novo mutations in epileptic encephalopathies. We bring statistical evidence that mutations in DNM1 cause epileptic encephalopathy, find suggestive evidence for a role of three additional genes, and show that at least 12% of analyzed individuals have an identifiable causal de novo mutation. Strikingly, 75% of mutations in these probands are predicted to disrupt a protein involved in regulating synaptic transmission, and there is a significant enrichment of de novo mutations in genes in this pathway in the entire cohort as well. These findings emphasize an important role for synaptic dysregulation in epileptic encephalopathies, above and beyond that caused by ion channel dysfunction.

The structure and conductivity of cerium and lanthanum phosphate glasses and glass-ceramics were investigated. The effects of varying the metal to phosphate ratio in the glasses, doping LaP3O9 glasses with Ce, and recrystallization of CeP3O9 glasses, on the glasses' microstructure and total conductivity were investigated using XRD, SEM, and AC impedance techniques. Strong increases in conductivity occurred when the glasses were recrystallized: the conductivity of a cerium metaphosphate glass increased conductivity after recrystallization from 10-7.5 S/cm to 10-6 S/cm at 400oC.

Nanoparticulate catalysts infiltrated into SOFC (Solid Oxide FUel Cell) electrodes can significantly enhance the cell performance, but the stability of these electrodes has been an open issue. An infiltration procedure is reported that leads to a stable scandia-stablized zirconia (SSZ) cathode electrode performance.

To improve the LSM-YSZ cathode performance of intermediate temperature solid oxide fuel cells (SOFCs), Sm0.6Sr0.4CoO3-sigma (SSC) perovskite nanoparticles are incorporated into the cathodes by a reaction-infiltration process. The SSC particles are ~;20 to 80nm in diameter, and intimately adhere to the pore walls of the preformed LSM-YSZ cathodes. The SSC particles dramatically enhance single-cell performance with a 97 percentH2+3 percentH2O fuel, between 600oC and 800oC. Consideration of a simplified TPB (triple phase boundary) reaction geometry indicates that the enhancement may be attributed to the high electrocatalytic activity of SSC for electrochemical reduction of oxygen in a region that can be located a small distance away from the strict triple phase boundaries. The implication of this work for developing high-performance electrodes is also d iscussed.

The fabrication and electrochemical performance of co-fired, LSM-SYSZ [i.e., La0.65Sr0.30MnO3 (LSM) - (Sc2O3)0.1(Y2O3)0.01(ZrO2)0.89] supported thin-film cells were examined using humidified hydrogen as a fuel. Co-firing of bi-layers and tri-layers was successful at 1250oC by optimizing the amount of carbon pore formers. A power density of a factor of 2.5 higher than that recently reported for the same type of cell at 800oC [3] was obtained for a cell with cobalt infiltration into the supporting cathode: the peak power densities were 455, 389, 285, 202, 141 mW/cm2 at 800, 750, 700, 650, 600oC, respectively, and in most cases power densities at 0.7V exceeded more than 90 percent of the peak output. Increasing the cathode porosity from 43 to 53 percent improved peak power densities by as much as 1.3, shifting the diffusion limitation to high current densities. Cobalt infiltration into the support improved those by as much as a factor of 2 due to a significant reduction in non-ohmic resistance. These results demonstrate that cobalt catalyst-infiltrated LSM can be effective and low-cost supporting electrodes for reduced temperature, thin film SOFCs.

An effective, dense and well adherent coating was produced on 430SS that has the result of significantly reducing the oxidation rate of this alloy at elevated temperatures. The coating is essentially a Mn-Co-O spinel, applied in powder form, and compacted to improve its green density. A simplified model is presented that allows an assessment of the effects of the contact and scale geometries. For 850oC, an ASR can be predicted of approximately 0.5 cm2, after 50,000hrs in air, taking into account a factor of 10 penalty for unfavorable contact geometries. The effect of the densified Mn-Co spinel coating is to reduce significantly Cr2O3 sub-scale formation, lower the thermal expansion mismatch, and increase the electronic conductivity of the scale. The findings point to several potential remedies for achieving coatings on 430 SS that allow for metal interconnects with a service life of 50,000hrs or more. Considering contact geometries, such service life is unlikely to be possible above operating temperatures of about 700oC, unless highly specialized alloys are used, with potential processing and cost penalties.

The effective oxygen conductivities in the zirconia networks of porous LSM-YSZ and LSM-SYSZ composites [i.e. La0.85Sr0.15MnO3 (Y2O3)0.08(ZrO2)0.92 and La0.85Sr0.15MnO3-c2O3)0.1(Y2O3)0.01(ZrO2)0.89, respectively] were evaluated by an AC impedance technique using specimens in which LSM was removed by hydrochloric acid leaching. The oxygen conductivities of porous YSZ and SYSZ alone followed a Koh-Fortini relationship. LSM-containing zirconia network conductivities were additionally decreased by the presence of the LSM, presumably by increased grain boundary resistances. Constriction resistances were estimated to have a minor effect.