Visualization and tracking of tubule-derived, fluorescent-labeled NS1 as a marker of bluetongue virus in living cells.

Bluetongue virus (BTV) is an arbovirus (genus, Orbivirus; family, Sedoreoviridae) transmitted among ruminant hosts by Culicoides biting midges (genus, Culicoides). Orbivirus sp. produce large numbers of virus-specific tubules during infection. Non-structural protein 1 (NS1) derived from BTV is a highly conserved tubule-forming protein that exists in infected cells as tubular and non-tubular forms. However, the lack of tagged viruses has limited dynamic tracking of NS1 tubule protein levels in living cells. Here, four recombinant viruses (BTV1-NS1-156TC, BTV1-NS1-156eGFP, BTV1-NS1-534eGFP, and BTV1-NS1-552eGFP) were successfully rescued using a BTV reverse genetics strategy by introduction of biarsenical-tetracysteine or enhanced green fluorescent protein (eGFP) tags to the BTV NS1 protein. Inserting these tags at these three sites resulted in different equilibrium states of NS1 between the tubular and non-tubular forms. NS1 was tracked in live cells to observe the dynamic movement and aggregation processes of NS1 tubules. The formation of NS1 tubules is a prerequisite for NS1 aggregation, and NS1 tubules are dependent on cellular microtubules to aggregate at the microtubule organizing center and encapsulate by vimentin to form aggresomes. This study confirms the possibility of controlling the conversion ratio between NS1 monomers and tubule forms, for the first time elucidates the movement and aggregation processes of NS1 tubules by live-cell imaging, and confirms that NS1 tubules form aggresomes. IMPORTANCE: While extensive research has established BTV as a model system to study large non-enveloped viruses, critical gaps remain in our understanding of its biology. During infection, BTV NS1 assembles into abundant tubulars, a hallmark feature of the Orbivirus genus. In this study, fluorescently tagged BTV NS1 proteins were engineered, and four recombinant viruses were successfully rescued. Insertion of these tags altered the equilibrium between tubular and monomeric NS1 conformations, demonstrating the feasibility of modulating the transition ratio between structural states. Additionally, NS1 tubules depend on microtubule-mediated intracellular transport for aggregation and subsequent aggresome formation. These findings provide a basis for the application of NS1 in vaccine delivery, therapy, nanotechnology, and future explorations of BTV pathogenesis

A new 3-arylbenzofuran derivative EIE-2 reestablishes Treg-dependent tolerance in rheumatoid arthritis by targeting on Syk induced mTOR and PKCθ imbalance

IntroductionDysfunctional self-tolerance is thought to play a crucial role in the onset of rheumatoid arthritis (RA) pathogenesis. Poorly functioning regulatory T cells (Tregs) lead to extreme situations where self-tolerance is robustly disrupted. However, there are many uncertainties regarding its immunosuppressive pathways, especially concerning therapeutic drugs that are still in their infancy. Therefore, deciphering potential targets and developing novel drugs to ameliorate functional Tregs deficiency appears to be an efficient therapeutic approach for controlling RA.MethodsThe therapeutic effects of EIE-2, a novel 3-arylbenzofuran derivative, were evaluated in collagen-induced arthritis (CIA) rats and carrageenan-induced paw edema mice in vivo, as well as in LPS-, PMA- or TNF-α-stimulated human CD4+ T cells (Jurkat), human synovial sarcoma cells (SW982) and primary isolated lymphocytes in vitro. The role of Syk in Treg-dependent tolerance and the mechanism of EIE-2 were explored using western blotting, quantitative reverse transcription PCR (qRT-PCR) and flow cytometry. Potential mechanistic targets were further validated through siRNA knockdown and molecular docking analysis.ResultsEIE-2 significantly ameliorated arthritic symptoms and pathological damage in CIA rats by reducing pro-inflammatory cytokines and increasing anti-inflammatory factors in synovium and serum, and exhibited similar therapeutic effects in carrageenan-induced pedal edema mice. Moreover, EIE-2 potently suppressed the inflammatory responses in human synoviocyte SW982 cells, primary isolated lymphocytes and CD4+ Jurkat cells. Its therapeutic potential was associated with upregulation of Tregs during the active phase and downregulation during the inactive phase of RA. Mechanistically, EIE-2 modulated the PKCθ/mTOR ratio via Syk targeting, thereby restoring homeostasis in Tregs.DiscussionEIE-2 is a potential therapeutic candidate for RA. The underlying mechanism may involve its targeting on Syk to upregulate the PKCθ/mTOR ratio during the active phase of RA and downregulate the PKCθ/mTOR ratio during the inactive phase of RA, ultimately promoting Treg-dependent tolerance restoration

QTL Mapping of Six Spike and Stem Traits in Hybrid Population of Agropyron Gaertn. in Multiple Environments

Most Agropyron Gaertn. species are excellent sources of forage. The derivative lines of wheat-Agropyron cristatum show elite agronomic traits, and some are valuable for wheat breeding. The species of Agropyron Gaertn. was mainly recognized by the spike morphology in traditional taxon. Six traits, including spike length (SL), ear stem length (ESL), the second internodes length (SIL), spikelet number per spike (SNS), floret number per spikelet (FNS), and grain number per spikelet (GNS), are vital to morphology studies and also influences the forage crop yield. To elucidate the genetic basis of spike and stem traits, a quantitative trait locus (QTL) analysis was conducted in a cross-pollinated (CP) hybrid population derived from a cross between two diverse parents, Agropyron mongolicum Keng Z2098 and A. cristatum (L.) Gaertn. Z1842, evaluated across three ecotopes (Langfang, Changli, and Guyuan of Hebei, China) over 3 years (from 2014 to 2016). Construction of a high-density linkage map was based on 1,023 single-nucleotide polymorphism (SNP) markers, covering 907.8 cM of the whole Agropyron genome. A total of 306 QTLs with single QTL in different environments explaining 0.07–33.21% of the phenotypic variation were detected for study traits. Seven major-effect QTLs were identified, including one for ESL on chromosome 3, one for SIL on chromosome 5, three for SL (two on chromosome 2 and one on chromosome 4), and two for SNS on chromosomes 3 and 7. Also, seven stable QTLs, including four for ESL, one for SL, one for GNS, and one for FNS, were mainly mapped on chromosomes 2, 3, 4, 5, and 7, respectively, elucidating 0.25–14.98% of the phenotypic variations. On the use of Agropyron CP hybrid population to identify QTL determining spike and stem traits for the first time, these QTLs for six traits would provide a theoretical reference for the molecular marker-assisted selection in the improvement of forage and cereal crop species

Regional Radiomics Similarity Networks Reveal Distinct Subtypes and Abnormality Patterns in Mild Cognitive Impairment.

Funder: Startup Funds for Leading Talents at Beijing Normal UniversityIndividuals with mild cognitive impairment (MCI) of different subtypes show distinct alterations in network patterns. The first aim of this study is to identify the subtypes of MCI by employing a regional radiomics similarity network (R2SN). The second aim is to characterize the abnormality patterns associated with the clinical manifestations of each subtype. An individual-level R2SN is constructed for N = 605 normal controls (NCs), N = 766 MCI patients, and N = 283 Alzheimer's disease (AD) patients. MCI patients' R2SN profiles are clustered into two subtypes using nonnegative matrix factorization. The patterns of brain alterations, gene expression, and the risk of cognitive decline in each subtype are evaluated. MCI patients are clustered into "similar to the pattern of NCs" (N-CI, N = 252) and "similar to the pattern of AD" (A-CI, N = 514) subgroups. Significant differences are observed between the subtypes with respect to the following: 1) clinical measures; 2) multimodal neuroimaging; 3) the proportion of progression to dementia (61.54% for A-CI and 21.77% for N-CI) within three years; 4) enriched genes for potassium-ion transport and synaptic transmission. Stratification into the two subtypes provides new insight for risk assessment and precise early intervention for MCI patients

CEP290 tethers flagellar transition zone microtubules to the membrane and regulates flagellar protein content

Entry and exit of proteins into flagella is gauged by CEP290 in the transition zone

Duckweed: a starch-hyperaccumulating plant under cultivation with a combination of nutrient limitation and elevated CO2

IntroductionThe increasing global demand for starch has created an urgent need to identify more efficient and sustainable production methods. However, traditional starch sources, such as crop-based options, experience significant bottlenecks due to limitations in land use, water consumption, and the impacts of climate change. Therefore, there is a pressing need to explore and develop new sources of starch.MethodsWe develop a novel duckweed cultivation technology that combines nutrients limitation and CO2 supplementation to achieve very high starch content. In this study, we integrated whole-genome sequencing, epigenomics, transcriptomics, enzyme activity, and composition variation to elucidate the mechanisms of efficient starch accumulation in duckweed in terms of starch accumulation and carbon partitioning, regulation of the expression of genes in the starch metabolic pathway, and sucrose biosynthesis and transportation.Results and discussionAlthough Landoltia punctata exhibits dramatic gene family contraction, its starch content and productivity reached 72.2% (dry basis) and 10.4 g m-2 d-1, respectively, in 10 days, equivalent to a yield of 38.0 t ha-1 y-1, under nutrient limitation treatment with elevated CO2 levels. We also examined the mechanism of high starch accumulation in duckweed. This phenomenon is associated with the regulation of DNA methylation and transcription factors as well as the significantly upregulated transcription levels and the increased activities of key enzymes involved in starch biosynthesis. Moreover, while nitrogen redistribution was increased, sucrose biosynthesis and transportation and lignocellulose biosynthesis were reduced. These alterations led to a reduction in lignocellulose and protein contents and ultimately an increase in the accumulation of starch in the chloroplasts.ConclusionThis work demonstrates the potential of duckweed as a highly efficient starch producer

The association of melanoma-associated antigen-A gene expression with clinicopathological characteristics and prognosis in resected non-small-cell lung cancer: a meta-analysis

Abstract Our goal was to explore the association of melanoma-associated antigen-A (MAGE-A) gene expression with clinicopathological parameters and survival rates in patients with non-small-cell lung cancer (NSCLC) who had surgery. A systematic search of EMBASE, PubMed, Web of Science and The Cochrane Library databases was performed through 20 April 2019. The combined risk ratios (RRs) and hazard ratios (HRs) with their corresponding 95% confidence intervals (CIs) were calculated to assess the association of MAGE-A gene expression with clinicopathological characteristics and prognosis of patients with resected NSCLC, respectively. All statistical analyses were performed with Stata software, version 12.0. A total of 9 articles involving 1538 patients were included in our meta-analysis; most of the studies were from Asian countries. The results indicated that the expression of the MAGE-A gene was significantly correlated with lymph node metastases (RR 1.21, 95% CI 1.09–1.34; P = 0.001), high tumour-node-metastasis stage (RR 1.24, 95% CI 1.12–1.38; P &lt; 0.001), histological type (squamous cell carcinoma) (RR 1.82, 95% CI 1.15–2.87; P = 0.01), poor overall survival (HR 2.11, 95% CI 1.73–2.57; P &lt; 0.001) and cancer-specific survival (HR 1.76, 95% CI 1.12–2.78; P = 0.015). MAGE-A gene expression is related to tumour development and metastasis and is more prevalent in squamous cell carcinomas of the lung; besides, it is an independent prognostic factor for patients with resected NSCLC.</jats:p