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Viruses often manipulate ubiquitination pathways to facilitate their replication and pathogenesis. CUL2ZYG11B known as the substrate receptor of cullin-2 RING E3 ligase, is bound by SARS-CoV-2 ORF10 to increase its E3 ligase activity, leading to degradation of IFT46, a protein component of the intraflagellar transport (IFT) complex B. This results in dysfunctional cilia, which explains certain symptoms that are specific to COVID-19. However, the precise molecular mechanism of how ORF10 recognizes CUL2ZYG11B remains unknown. Here, we determined the crystal structure of CUL2ZYG11B complexed with the N-terminal extension (NTE) of SARS-CoV-2 ORF10 (2.9 Å). The structure reveals that the ORF10 N-terminal heptapeptide (NTH) mimics the Gly/N-degron to bind CUL2ZYG11B. Mutagenesis studies identified key residues within ORF10 that are key players in its interaction with CUL2ZYG11B both in ITC assay and in vivo cells. In addition, we prove that enhancement of CUL2ZYG11B activity for IFT46 degradation by which ORF10-mediated correlates with the binding affinity between ORF10 and CUL2ZYG11B. Finally, we used a Global Protein Stability system to show that the NTH of ORF10 mimics the Gly/N-degron motif, thereby binding competitively to CUL2ZYG11B and inhibiting the degradation of target substrates bearing the Gly/N-degron motif. Overall, this study sheds light on how SARS-CoV-2 ORF10 exploits the ubiquitination machinery for proteasomal degradation, and offers valuable insights for optimizing PROTAC-based drug design based on NTH CUL2ZYG11B interaction, while pinpointing a promising target for the development of treatments for COVID-19.

Abstract SETD3 is an essential host factor for the replication of a variety of enteroviruses that specifically interacts with viral protease 2A. However, the interaction between SETD3 and the 2A protease has not been fully characterized. Here, we use X-ray crystallography and cryo-electron microscopy to determine the structures of SETD3 complexed with the 2A protease of EV71 to 3.5 Å and 3.1 Å resolution, respectively. We find that the 2A protease occupies the V-shaped central cleft of SETD3 through two discrete sites. The relative positions of the two proteins vary in the crystal and cryo-EM structures, showing dynamic binding. A biolayer interferometry assay shows that the EV71 2A protease outcompetes actin for SETD3 binding. We identify key 2A residues involved in SETD3 binding and demonstrate that 2A’s ability to bind SETD3 correlates with EV71 production in cells. Coimmunoprecipitation experiments in EV71 infected and 2A expressing cells indicate that 2A interferes with the SETD3-actin complex, and the disruption of this complex reduces enterovirus replication. Together, these results reveal the molecular mechanism underlying the interplay between SETD3, actin, and viral 2A during virus replication.

Summary: Inhibition of nucleic acid targets is mediated by Argonaute (Ago) proteins guided by RNA or DNA. Although the mechanisms underpinning the functions of eukaryotic and “long” prokaryotic Ago proteins (pAgos) are well understood, those for short pAgos remain enigmatic. Here, we determine two cryoelectron microscopy structures of short pAgos in association with the NADase-domain-containing protein Sir2-APAZ from Geobacter sulfurreducens (GsSir2/Ago): the guide RNA-target DNA-loaded GsSir2/Ago quaternary complex (2.58 Å) and the dimer of the quaternary complex (2.93Å). These structures show that the nucleic acid binding causes profound conformational changes that result in disorder or partial dissociation of the Sir2 domain, suggesting that it adopts a NADase-active conformation. Subsequently, two RNA-/DNA-loaded GsSir2/Ago complexes form a dimer through their MID domains, further enhancing NADase activity through synergistic effects. The findings provide a structural basis for short-pAgo-mediated defense against invading nucleic acids.

Abstract Microbial species often occur in complex communities and exhibit intricate synergistic and antagonistic interactions. To avoid predation and compete for favorable niches, bacteria have evolved specialized protein secretion systems. The type VI secretion system (T6SS) is a versatile secretion system widely distributed among Gram-negative bacteria that translocates effectors into target cells or the extracellular milieu via various physiological processes. Pseudomonas aeruginosa is an opportunistic pathogen responsible for many diseases, and it has three independent T6SSs (H1-, H2-, and H3-T6SS). In this study, we found that the H3-T6SS of highly virulent P. aeruginosa PA14 is negatively regulated by OxyR and OmpR, which are global regulatory proteins of bacterial oxidative and acid stress. In addition, we identified a H3-T6SS effector PA14_33970, which is located upstream of VgrG3. PA14_33970 interacted directly with VgrG3 and translocated into host cells. Moreover, we found that H3-T6SS and PA14_33970 play crucial roles in oxidative, acid, and osmotic stress resistance, as well as in motility and biofilm formation. PA14_33970 was identified as a new T6SS effector promoting biofilm formation and thus named TepB. Furthermore, we found that TepB contributes to the virulence of P. aeruginosa PA14 toward Caenorhabditis elegans. Overall, our study indicates that H3-T6SS and its biofilm-promoting effector TepB are regulated by OxyR and OmpR, both of which are important for adaptation of P. aeruginosa PA14 to multiple stressors, providing insights into the regulatory mechanisms and roles of T6SSs in P. aeruginosa.

Sarbecovirus ORF6 binds to the Rae1-Nup98 complex, a component of the cytoplasmic face of the nuclear pore complex, and has been shown to suppress interferon responses. Here, the authors provide structures of Rae1-Nup98 in complex with the C-terminal tails of SARS-CoV-2 and SARS-CoV ORF6 and provide insights into ORF6-mediated blockade of mRNA and STAT1 nucleocytoplasmic transport.

SARS-CoV-2 orf9b binds to the mitochondrial outer membrane protein TOM70 and has been linked to the suppression of interferon responses. Here, the authors characterize the interactions of SARS-CoV-2 orf9b and human TOM70 biochemically, and they determine the 2.2 Å crystal structure of the TOM70 cytosolic domain with a bound SARS-CoV-2 orf9b peptide.

The pandemic of coronavirus disease 2019 (COVID-19) is changing the world like never before. This crisis is unlikely contained in the absence of effective therapeutics or vaccine. The papain-like protease (PLpro) of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) plays essential roles in virus replication and immune evasion, presenting a charming drug target. Given the PLpro proteases of SARS-CoV-2 and SARS-CoV share significant homology, inhibitor developed for SARS-CoV PLpro is a promising starting point of therapeutic development. In this study, we sought to provide structural frameworks for PLpro inhibitor design. We determined the unliganded structure of SARS-CoV-2 PLpro mutant C111S, which shares many structural features of SARS-CoV PLpro. This crystal form has unique packing, high solvent content and reasonable resolution 2.5 Å, hence provides a good possibility for fragment-based screening using crystallographic approach. We characterized the protease activity of PLpro in cleaving synthetic peptide harboring nsp2/nsp3 juncture. We demonstrate that a potent SARS-CoV PLpro inhibitor GRL0617 is highly effective in inhibiting protease activity of SARS-CoV-2 with the IC50 of 2.2 ± 0.3 μmol/L. We then determined the structure of SARS-CoV-2 PLpro complexed by GRL0617 to 2.6 Å, showing the inhibitor accommodates the S3–S4 pockets of the substrate binding cleft. The binding of GRL0617 induces closure of the BL2 loop and narrows the substrate binding cleft, whereas the binding of a tetrapeptide substrate enlarges the cleft. Hence, our results suggest a mechanism of GRL0617 inhibition, that GRL0617 not only occupies the substrate pockets, but also seals the entrance to the substrate binding cleft hence prevents the binding of the LXGG motif of the substrate.

Background and Objective:Parkinson’s disease (PD) is a common neurodegenerative disease. Previous studies have demonstrated the effect of cell-based therapies, but their clinical efficacy and safety have not been evaluated. This review protocol aimed to systematically evaluate the effect of stem cell therapy in patients with PD and to develop an evidence base for guiding policy and practice.Methods:PubMed, Embase, MedlinePlus, The Lancet and Brain were searched over the period January 2001 to October 2019. The keywords used for searching were "Parkinson’s disease" and "cell therapy" and "mesenchymal stem cells" and "embryonic stem cells" and "brain-derived neural stem cells" and "neural progenitor cells" . The Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) statement and a measurement tool, Assessment of Multiple Systematic Reviews (AMSTAR), to assess systematic reviews were used to assess the reporting quality and methodological quality. Data extracted included study details, participant details, intervention details and outcome.Results:Nine valid research papers were screened out by systematic analysis. These nine studies were carried out in different countries, with different populations and cell types. According to evaluation methods used, all of the transplantation therapies reported can improve the symptoms of PD patients.Conclusions:Cell transplantation is a potential treatment option for PD. More studies with strict study design, larger sample sizes, and longer follow-up are needed in the future.

Alongshan virus (ALSV) is an emerging human pathogen that was identified in China and rapidly spread to the European continent in 2019, raising concerns about public health. ALSV belongs to the distinct Jingmenvirus group within the Flaviviridae family with segmented RNA genomes. While segments 2 and 4 of the ALSV genome encode the VP1–VP3 proteins of unknown origin, segments 1 and 3 encode the NS2b–NS3 and NS5 proteins, which are related to Flavivirus nonstructural proteins, suggesting an evolutionary link between segmented and unsegmented viruses within the Flaviviridae family. Here, the enzymatic activity of the ALSV NS3-like helicase (NS3-Hel) was characterized and its crystal structure was determined to 2.9 Å resolution. ALSV NS3-Hel exhibits an ATPase activity that is comparable to those measured for Flavivirus NS3 helicases. The structure of ALSV NS3-Hel exhibits an overall fold similar to those of Flavivirus NS3 helicases. Despite the limited amino-acid sequence identity between ALSV NS3-Hel and Flavivirus NS3 helicases, structural features at the ATPase active site and the RNA-binding groove remain conserved in ALSV NS3-Hel. These findings provide a structural framework for drug design and suggest the possibility of developing a broad-spectrum antiviral drug against both Flavivirus and Jingmenvirus.

Pseudomonas aeruginosa (P. aeruginosa) is a typical Gram-negative bacterium that can cause the spoilage of catered food products. Using a luminescent reporter gene (luxCDABE), this study sought to construct a cell-based biosensor (PAO1-CE) to rapidly screen antibacterial substances against P. aeruginosa. A total of six antibiotics belonging to five categories were used as the model test substances. The results of the bioluminescence detection method were verified using traditional antibacterial research assessments. The correlation coefficient of the regression equation fitting the data generated using this method was greater than 0.98, supporting the credibility of this approach. Additionally, the EC50 of each of the antibiotics assessed in this study was lower than the 1/2 MIC determined by conventional means. All six of the antibiotics caused varying degrees of damage to the cell membrane and cell wall of P. aeruginosa. Importantly, this novel method helped shorten the time necessary for active-compound detection and could be used for high-throughput detection, which would also help improve the detection efficiency. The application of this method towards the discovery of novel antibacterial compounds targeting P. aeruginosa holds substantial promise for greatly improving the efficiency of compound discovery.

Mycobacterium tuberculosis (Mtb) caused an estimated 10 million cases of tuberculosis and 1.2 million deaths in 2019 globally. The increasing emergence of multidrug-resistant and extensively drug-resistant Mtb is becoming a public health threat worldwide and makes the identification of anti-Mtb drug targets urgent. Elongation factor G (EF-G) is involved in tRNA translocation on ribosomes during protein translation. Therefore, EF-G is a major focus of structural analysis and a valuable drug target of antibiotics. However, the crystal structure of Mtb EF-G1 is not yet available, and this has limited the design of inhibitors. Here, we report the crystal structure of Mtb EF-G1 in complex with GDP. The unique crystal form of the Mtb EF-G1-GDP complex provides an excellent platform for fragment-based screening using a crystallographic approach. Our findings provide a structure-based explanation for GDP recognition, and facilitate the identification of EF-G1 inhibitors with potential interest in the context of drug discovery.

Although educational timetabling problems have been studied for decades, one instance of this, the school timetabling problem (STP), has not developed as quickly as examination timetabling and course timetabling problems due to its diversity and complexity. In addition, most STP research has only focused on the educators’ availabilities when studying the educator aspect, and the educators’ preferences and expertise have not been taken into consideration. To fill in this gap, this paper proposes a conceptual model for the school timetabling problem considering educators’ availabilities, preferences and expertise as a whole. Based on a common real-world school timetabling scenario, the artificial bee colony (ABC) algorithm is adapted to this study, as research shows its applicability in solving examination and course timetabling problems. A virtual search space for dealing with the large search space is introduced to the proposed model. The proposed approach is simulated with a large, randomly generated dataset. The experimental results demonstrate that the proposed approach is able to solve the STP and handle a large dataset in an ordinary computing hardware environment, which significantly reduces computational costs. Compared to the traditional constraint programming method, the proposed approach is more effective and can provide more satisfactory solutions by considering educators’ availabilities, preferences, and expertise levels.

Pathogens require iron for their metabolism and virulence. Here the authors identify an iron acquisition system inPseudomonas aeruginosainvolving a protein secreted by a type VI secretion system, the PQS signalling compound and siderophore receptors.

Urease acts as an important acid resistance system and virulence factor that is widespread among microorganisms. RovM is a global regulator that regulates a series of genes and pathways including acid survival systems in the enteric bacterium Yersinia pseudotuberculosis (Yptb). However, whether RovM regulates the urease activity in Yptb was still unknown. In this study, by using qualitative and quantitative urease assays, we show that the urease expression responds to nutrient conditions and the RovM protein represses urease expression by binding to its promoter. A previously reported positive regulator OmpR activates urease activity but RovM plays a dominant role in different nutrient conditions. In addition, carbon storage regulator system A (CsrA), the upstream regulator of RovM, dramatically down-regulates urease activity possibly by its binding to the Shine-Dalgarno (SD) sequence of the mRNA encoding the urease. In conclusion, this study demonstrates that urease activity is strictly controlled by nutrient conditions and is down-regulated by the CsrA-RovM pathway.

Abstract Background Hypoderaeum conoideum is a neglected but important trematode. The life cycle of this parasite is complex: snails serve as the first intermediate hosts: bivalves, fishes or tadpoles serve as the second intermediate hosts, and poultry (such as chickens and ducks) act as definitive hosts. In recent years, H. conoideum has caused significant economic losses to the poultry industry in some Asian countries. Despite its importance, little is known about the molecular ecology and population genetics of this parasite. Knowledge of mitochondrial (mt) genome of H. conoideum can provide a foundation for phylogenetic studies as well as epidemiological investigations. Methods The entire mt genome of H. conoideum was amplified in five overlapping fragments by PCR and sequenced, annotated and compared with mt genomes of selected trematodes. A phylogenetic analysis of concatenated mt amino acid sequence data for H. conoideum, eight other digeneans (Clonorchis sinensis, Fasciola gigantica, F. hepatica, Opisthorchis felineus, Schistosoma haematobium, S. japonicum, S. mekongi and S. spindale) and one tapeworm (Taenia solium; outgroup) was conducted to assess their relationships. Results The complete mt genome of H. conoideum is 14,180 bp in length, and contains 12 protein-coding genes, 22 transfer RNA genes, two ribosomal RNA genes and one non-coding region (NCR). The gene arrangement is the same as in Fasciola spp, with all genes being transcribed in the same direction. The phylogenetic analysis showed that H. conoideum had a relatively close relationship with F. hepatica and other members of the Fasciolidae, followed by the Opisthorchiidae, and then the Schistosomatidae. Conclusions The mt genome of H. conoideum should be useful as a resource for comparative mt genomic studies of trematodes and for DNA markers for systematic, population genetic and epidemiological studies of H. conoideum and congeners.

Abstract Creativity is typically operationalized as divergent thinking (DT) ability, a form of higher-order cognition which relies on memory, attention, and other component processes. Despite recent advances, creativity neuroscience lacks a unified framework to model its complexity across neural, genetic, and cognitive scales. Using task-based fMRI from two independent samples and MVPA, we identified a neural pattern that predicts DT, validated through cognitive decoding, genetic data, and large-scale resting-state fMRI. Our findings reveal that DT neural patterns span brain regions associated with diverse cognitive functions, with positive weights in the default mode and frontoparietal control networks and negative weights in the visual network. The high correlation with the primary gradient of functional connectivity suggests that DT involves extensive integration from concrete sensory information to abstract, higher-level cognition, distinguishing it from other advanced cognitive functions. Moreover, neurobiological analyses show that the DT pattern is positively correlated with dopamine-related neurotransmitters and genes influencing neurotransmitter release, advancing the neurobiological understanding of creativity.

Abstract Novelty and appropriateness are two fundamental components of creativity. However, the way in which novelty and appropriateness are separated at behavioral and neural levels remains poorly understood. In the present study, we aim to distinguish behavioral and neural bases of novelty and appropriateness of creative idea generation. In alignment with two established theories of creative thinking, which respectively, emphasize semantic association and executive control, behavioral results indicate that novelty relies more on associative abilities, while appropriateness relies more on executive functions. Next, employing a connectome predictive modeling (CPM) approach in resting-state fMRI data, we define two functional network-based models—dominated by interactions within the default network and by interactions within the limbic network—that respectively, predict novelty and appropriateness (i.e., cross-brain prediction). Furthermore, the generalizability and specificity of the two functional connectivity patterns are verified in additional resting-state fMRI and task fMRI. Finally, the two functional connectivity patterns, respectively mediate the relationship between semantic association/executive control and novelty/appropriateness. These findings provide global and predictive distinctions between novelty and appropriateness in creative idea generation.

Diseases caused by parasitic helminths cause considerable production and economic losses in livestock worldwide. Understanding the epidemiology of these parasites has important implications for controlling them. The main purpose of the present study was to estimate the prevalence of key parasitic helminths in goats along the Han River in Zhanggang, Hubei Province (from January to December 2014). We used faecal flotation and sedimentation techniques as well as PCR-based DNA sequencing to detect and identify helminths. Results showed that the prevalence of helminths was high throughout the year, particularly for gastrointestinal nematodes. These first findings provide useful baseline information for goat helminths in Zhanggang, and a starting point for the implementation of control programs. With an increased expansion of the goat industry in China, the findings also emphasise the need to undertake prevalence surveys in other regions of China where extensive farming practices are used. [ABSTRACT FROM AUTHOR]

Although many studies have explored the neural mechanism of the feeling of stress, to date, no effort has been made to establish a model capable of predicting the feeling of stress at the individual level using the resting-state functional connectome. Although individuals may be confronted with multidimensional stressors during the coronavirus disease 2019 (COVID-19) pandemic, their appraisal of the impact and severity of these events might vary. In this study, connectome-based predictive modeling (CPM) with leave-one-out cross-validation was conducted to predict individual perceived stress (PS) from whole-brain functional connectivity data from 817 participants. The results showed that the feeling of stress could be predicted by the interaction between the default model network and salience network, which are involved in emotion regulation and salience attribution, respectively. Key nodes that contributed to the prediction model comprised regions mainly located in the limbic systems and temporal lobe. Critically, the CPM model of PS based on regular days can be generalized to predict individual PS levels during the COVID-19 pandemic, which is a multidimensional, uncontrollable stressful situation. The stability of the results was demonstrated by two independent datasets. The present work not only expands existing knowledge regarding the neural mechanism of PS but also may help identify high-risk individuals in healthy populations.

Creative thinking is a hallmark of human cognition, which enables us to generate novel and useful ideas. Nevertheless, its emergence within the macro-scale neurocognitive circuitry remains largely unknown. Using resting-state fMRI data from two large population samples (SWU: n = 931; HCP: n = 1001) and a novel “travelling pattern prediction analysis”, here we identified the modularized functional connectivity patterns linked to creative thinking ability, which concurrently explained individual variability across ordinary cognitive abilities such as episodic memory, working memory and relational processing. Further interrogation of this neural pattern with graph theoretical tools revealed both hub-like brain structures and globally-efficient information transfer paths that together may facilitate higher creative thinking ability through the convergence of distinct cognitive operations. Collectively, our results provide reliable evidence for the hypothesized emergence of creative thinking from core cognitive components through neural integration, and thus allude to a significant theoretical advancement in the study of creativity.

Stress-related psychosomatic responses are viewed as important risks to our physical health. Growing evidence from structural imaging studies has implicated that stress and trauma exposures have negative effects on brain structural alterations. However, whether stress-related emotional distress and somatic symptoms are related to the structure of brain systems remains unclear. Also, stress-related somatic symptoms have adverse effects on emotional distress. In turn, emotional distress may influence somatic symptom reports via negative cognitive bias. However, whether this relationship is mediated by specific brain morphology remains poorly understood. First, we used voxel-based morphometric approaches to investigate the neuroanatomical basis underlying somatic symptoms and emotional distress in a large sample of healthy subjects (ages 18–27 years). We found that relatively high stress-related somatic symptoms were associated with reduced gray matter volumes (GMVs) in the ventral medial prefrontal cortex (vmPFC), anterior insula, somatosensory cortex, hippocampus, and amygdala. Furthermore, a moderator analysis was performed to investigate the impact of recent stressful life events (moderators) on the association between specific GMVs (independent variables) and emotional distress (dependent variables). Interestingly, high levels of emotional distress were associated with small volumes of the vmPFC, anterior insula, hippocampus, and amygdala in participants with experience with more recent stressful life events. Finally, we performed mediation analyses to investigate the specific brain areas that mediate the association between emotional distress and somatic symptoms. The results showed that the effect of emotional distress on somatic symptoms is mediated by reductions in the volume of the hippocampus, the impact of somatic symptoms on emotional distress is mediated by the volume of the vmPFC. These results provided evidence that higher stress-related somatic symptoms are associated with smaller volume in prefrontal, insula, and limbic regions involved in emotion, interoception, and memory processing. The vmPFC and hippocampus play different roles in the relationship between emotional distress and somatic symptoms.

Abstract The most widely used measurements of mindfulness are the Mindful Attention Awareness Scale (MAAS) and the Five Facet Mindfulness Questionnaire (FFMQ). However, controversies exist regarding the application of these scales. Additionally, the neural mechanisms of dispositional mindfulness havebecome a topic of interest. In the current study, we used surface-based methodology to identify the brain regions underlying individual differences in dispositional mindfulness in a large non-clinical sampleand compared the two instruments for measuring the dispositional mindfulness. The results indicated that theMAAS scores were significantly associated with increased greymatter volumes in the right precuneus and the significant association between the precuneus and depression symptomatology was mediated by MAAS scores. Regarding the FFMQ, the Describing, Nonjudging, and Nonreactivity facets were selectively associated with the cortical volume, thickness and surface area of multiple prefrontal regions as well as the inferior parietal lobule. Importantly, Describing mediated the association between the dorsolateral PFC volume and the cognitive reappraisal strategies of emotion regulation. These resultssuggested that the MAASwere mainly associated with self-awareness, while the FFMQ facets were selectively involved in emotion regulation, attention control and self-awareness. Therefore, this study characterized the differences in inter-individual variability between the two typical measurements of dispositional mindfulnessand the correlations between those measurements and imaging analyses.

Little is known about the association between emotional intelligence (EI) and trait creativity (TC), and the brain structural bases which involves. This study investigated the neuroanatomical basis of the association between EI and TC which measured by the Schutte self-report EI scale and the Williams creativity aptitude test. First, the voxel-based morphometry (VBM) analysis was used to explore the brain structures which is closely related to EI in a large young sample (n = 213). The results showed that EI was positively correlated with the regional gray matter volume (rGMV) in the right orbitofrontal cortex (OFC), which is regarded as a key region of emotional processing. More importantly, further mediation analysis revealed that rGMV in the right OFC partially mediated the association between EI and TC, which showed the OFC volume could account for the relationship between EI and TC. These findings confirmed the close relationship between EI and TC, and highlighted that the brain volumetric variation in the OFC associated with the top-down processing of emotion regulation, which may play a critical role in the promotion of TC. Together, these findings contributed to sharpening the understanding of the complex relationship between EI and TC from the perspective of brain structural basis.