Investigating the Evolution of Drosophila STING-Dependent Antiviral Innate Immunity by Multispecies Comparison of 2'3'-cGAMP Responses.

Viruses represent a major threat to all animals, which defend themselves through induction of a large set of virus-stimulated genes that collectively control the infection. In vertebrates, these genes include interferons that play a critical role in the amplification of the response to infection. Virus- and interferon-stimulated genes include restriction factors targeting the different steps of the viral replication cycle, in addition to molecules associated with inflammation and adaptive immunity. Predictably, antiviral genes evolve dynamically in response to viral pressure. As a result, each animal has a unique arsenal of antiviral genes. Here, we exploit the capacity to experimentally activate the evolutionarily conserved stimulator of IFN genes (STING) signaling pathway by injection of the cyclic dinucleotide 2'3'-cyclic guanosine monophosphate-adenosine monophosphate into flies to define the repertoire of STING-regulated genes in 10 Drosophila species, spanning 40 million years of evolution. Our data reveal a set of conserved STING-regulated factors, including STING itself, a cGAS-like-receptor, the restriction factor pastel, and the antiviral protein Vago, but also 2 key components of the antiviral RNA interference pathway, Dicer-2, and Argonaute2. In addition, we identify unknown species- or lineage-specific genes that have not been previously associated with resistance to viruses. Our data provide insight into the core antiviral response in Drosophila flies and pave the way for the characterization of previously unknown antiviral effectors.

Identification of Increased Grain Length 1 (IGL1), a novel gene encoded by a major QTL for modulating grain length in rice.

KEY MESSAGE: A novel quantitative trait locus qIGL1, which performed a positive function in regulating grain length in rice, was cloned by the map-based cloning approach; further studies revealed that it corresponded to LOC_Os03g30530, and the IGL1 appeared to contribute to lengthening and widening of the cells on the surface of grain hulls. Grain length is a prominent determinant for grain weight and appearance quality of rice. In this study, we conducted quantitative trait locus mapping to determine a genomic interval responsible for a long-grain phenotype observed in a japonica cultivar HD385. This led to the identification of a novel QTL for grain length on chromosome 3, named qIGL1 (for Increased Grain Length 1); the HD385 (Handao 385)-derived allele showed enhancement effects on grain length, and such an allele as well as NIP (Nipponbare)-derived allele was designated qigl1 HD385 and qIGL1NIP, respectively. Genetic analysis revealed that the qigl1HD385 allele displayed semidominant effects on grain length. Fine mapping further narrowed down the qIGL1 to an ~ 70.8-kb region containing 9 open reading frames (ORFs). A comprehensive analysis indicated that LOC_Os03g30530, which corresponded to ORF6 and carried base substitutions and deletions in HD385 relative to NIP, thereby causing changes or losses of amino-acid residues, was the true gene for qIGL1. Comparison of grain traits between a pair of near-isogenic lines (NILs), termed NIL-igl1HD385 and NIL-IGL1NIP, discovered that introduction of the igl1HD385 into the NIP background significantly resulted in the elevations of grain length and 1000-grain weight. Closer inspection of grain surfaces revealed that the cell length and width in the longitudinal direction were significantly longer and greater, respectively, in NIL-igl1HD385 line compared with in NIL-IGL1NIP line. Hence, our studies identified a new semidominant natural allele contributing to the increase of grain length and further shed light on the regulatory mechanisms of grain length.

The virus-induced cyclic dinucleotide 2'3'-c-di-GMP mediates STING-dependent antiviral immunity in Drosophila.

In mammals, the enzyme cGAS senses the presence of cytosolic DNA and synthesizes the cyclic dinucleotide (CDN) 2'3'-cGAMP, which triggers STING-dependent immunity. In Drosophila melanogaster, two cGAS-like receptors (cGLRs) produce 3'2'-cGAMP and 2'3'-cGAMP to activate STING. We explored CDN-mediated immunity in 14 Drosophila species covering 50 million years of evolution and found that 2'3'-cGAMP and 3'2'-cGAMP failed to control infection by Drosophila C virus in D. serrata and two other species. We discovered diverse CDNs produced in a cGLR-dependent manner in response to viral infection in D. melanogaster, including 2'3'-c-di-GMP. This CDN was a more potent STING agonist than cGAMP in D. melanogaster and it also activated a strong antiviral transcriptional response in D. serrata. Our results shed light on the evolution of cGLRs in flies and provide a basis for understanding the function and regulation of this emerging family of pattern recognition receptors in animal innate immunity.

A novel virus-induced cyclic dinucleotide, 2'3'-c-di-GMP, mediates STING-dependent antiviral immunity in Drosophila.

In mammals, the enzyme cGAS senses the presence of cytosolic DNA and synthesizes the cyclic dinucleotide (CDN) 2'3'-cGAMP. This CDN binds to and activates the protein STING to trigger immunity. We recently discovered in the model organism Drosophila melanogaster two cGAS-like receptors (cGLRs) that activate STING-dependent antiviral immunity and can produce 3'2'-cGAMP, in addition to 2'3'-cGAMP. Here we explore CDN-mediated immunity in 14 different Drosophila species covering 50 million years of evolution and report that 2'3'-cGAMP and 3'2'-cGAMP fail to control infection by Drosophila C virus in D. serrata, D. sechellia and D. mojavensis . Using an accurate and sensitive mass spectrometry method, we discover an unexpected diversity of CDNs produced in a cGLR-dependent manner in response to viral infection in D. melanogaster , including a novel CDN, 2'3'-c-di-GMP. We show that 2'3'-c-di-GMP is the most potent STING agonist identified so far in D. melanogaster and that this molecule also activates a strong antiviral transcriptional response in D. serrata . Our results shed light on the evolution of cGLRs in flies and provide a basis for the understanding of the function and regulation of this emerging family of PRRs in animal innate immunity.

Hematodinium perezi naturally infects Asian brush-clawed crab (Hemigrapsus takanoi).

The parasitic dinoflagellates of the genus Hematodinium have been considered one of the most important emerging pathogens for a broad range of marine crustaceans around the world. In China, frequent outbreaks of Hematodinium infections have caused serious economic losses for local farmers since 2004. Wild crabs were recently indicated to play a vital role in the transmission and spreading of the Hematodinium disease in polyculture pond systems. Based on PCR amplification and histopathological examination, we demonstrated that H. perezi can naturally infect a wild crab species, Hemigrapsus takanoi, which were collected from the waterways located on the coast of Rizhao or Weifang, Shandong Peninsula, China. According to the sequence similarity analysis and phylogenetic analysis, the Hematodinium isolates were identified as H. perezi and belonged to genotype II. The prevalence of H. perezi ranged from 3.3% to 5.7% in H. takanoi originating from Rizhao (n = 165 wild crabs) and from 0.9% to 20.0% in that originating from Weifang (n = 1386 wild crabs), respectively. To our knowledge, H. takanoi is, for the first time, reported as a new host for Hematodinium. Given the wide distribution of H. takanoi on the coasts along the Shandong Peninsula and the relative high prevalence of infection we monitored in our study, we speculate that H. takanoi contributes to the introducing and spreading parasitic Hematodinium between ponds via waterways in a poly-culturing system. Findings in this study broaden the host range of this parasite and expand the scope of our surveillance for Hematodinium disease in China.

Explainable machine learning-assisted origin identification: Chemical profiling of five lotus (Nelumbo nucifera Gaertn.) parts.

Five homologous lotus parts, namely, the leaf, stamen, plumule, flower and leaf base, are all ancient nutrient sources, but their chemical differences are poorly understood. Identification of these parts of origin could contribute to determining reasonable edible and/or medicinal applications without misuse/waste risk. The present work aimed to investigate the feasibility of using metabolic profiles coupled with explainable machine learning (ML) for tracing lotus parts of origin. Assisted with molecular networking, 151 compounds were systematically annotated through an untargeted metabolomics approach. Twenty-eight representative constituents were subsequently quantified for the construction of the ML algorithm. Because most ML algorithms are data-driven black boxes with opaque inner workings, the SHaply Additive exPlanation technique was innovatively used to understand model outputs. By offering an integral analytical platform for phytochemical characterization and information interpretation, these results could serve as a basis for an explainable tool for identification of the specific lotus part of origin.

Factors Associated with the Uptake of Genetic Testing for Cancer Risks: A Pathway Analysis Using the Health Information National Trends Survey Data.

Our study aimed to identify pathways from the source of information to the uptake of cancer genetic testing, with consideration of intermediate variables including perceptional, attitudinal and psychosocial factors. We used the Health Information National Trends Survey (2020 database) and constructed a structural equation model for pathway analysis (using SPSS version 24). Variables for socio-demographic, lifestyle and health information were also collected and used for confounding adjustment. A total of 2941 participants were analyzed (68.5%, non-Hispanic white; 59.7%, females; 58 years, median age; and 142 (4.8%) had undertaken genetic testing for cancer risk previously). Our pathway analysis found that only information from particular sources (i.e., healthcare providers and genetic counsellors) had positive and significant effects on people's perceptions of cancer regarding its prevention, detection and treatment (standardized ß range, 0.15−0.31, all p-values < 0.01). Following the paths, these perceptional variables (cancer prevention, detection and treatment) showed considerable positive impacts on the uptake of genetic testing (standardized ß (95% CIs): 0.25 (0.20, 0.30), 0.28 (0.23, 0.33) and 0.12 (0.06, 0.17), respectively). Pathways involving attitudinal and psychosocial factors showed much smaller or insignificant effects on the uptake of genetic testing. Our study brings several novel perspectives to the behavior model and may underpin certain issues regarding cancer risk genetic testing.

LncRNA-mRNA integrated profiling analysis in response to white spot syndrome virus in hepatopancreas in Penaeus japonicus.

Penaeus japonicas is an important shrimp species, which is exposed to stressors including a variety of epidemic diseases. To date, little is known about the mechanisms involved in the response to white spot syndrome virus (WSSV) mediated by long non-coding RNAs (lncRNAs). A total of 6544 putative lncRNAs were identified in the hepatopancreas in P. japonicas, which provides a useful lncRNA reference resource for use in future studies. In addition, a total of 444 differentially expressed mRNAs and 457 differentially expressed lncRNAs were identified at 6, 12, and 24 h after WSSV infection in the hepatopancreas of P. japonicas. Functional enrichment analysis showed that the differentially expressed mRNAs were enriched in terms related to immune response and viral infectivity such as defense response, aminopeptidase activity, whereas the differentially expressed lncRNA partner genes were enriched in ubiquitin-dependent protein catabolic process, lipoprotein metabolic process, and antigen processing and presentation. Moreover, several lncRNAs were induced by WSSV infection, indicating these lncRNAs might participate in regulating many immune processes referring to their partner genes. Co-expression analysis of the lncRNAs and their partner genes identified some high lncRNA-mRNA correlations. These results suggest that WSSV stimulates the immune response in the hepatopancreas potentially through an important coding and non-coding gene network, thereby providing valuable information regarding non-coding responses to WSSV in Penaeus species.

Oxidizing Hexagonal Boron Nitride into Fluorescent Structures by Photodissociated Directional Oxygen Radical.

Modifying the wide band gap semiconductor hexagonal boron nitride (hBN) can bring new chances in photonics. By virtue of the solvothermal/hydrothermal oxidation or functionalization, hBN can be converted into fluorescent nanodots. Until now, it has been a big challenge to drily oxidize hBN and turn it into bright fluorescent structures due to its superior chemical stability. Here, we report the oxidation of multilayer hBN into fluorescent structures by ultraviolet (UV, λ = 172 nm) photodissociated directional oxygen radical [O(3P)] in a gradient magnetic field. The paramagnetic O(3P), produced in a low-pressure O2 atmosphere, drifts toward hBN and then converts it into boron nitride oxide (BNO) micro/nanometer structures constituted by BO, BO2, and O-doped hBN. For a properly oxidized BNO substance, bright and photostable wide-band photoluminescence is realized with nanosecond-scaled lifetimes under the excitation of UV and visible lights.

Remineralization of human dentin type I collagen fibrils induced by carboxylated polyamidoamine dendrimer/amorphous calcium phosphate nanocomposite: an in vitro study.

Intrafibrillar mineralization of type I collagen fibrils is of great significance in dental remineralization, which is the key of caries prevention and treatment. Herein, two substances that have the remineralization ability, carboxylated polyamidoamine dendrimer (PAMAM-COOH) and nano-sized amorphous calcium phosphate (n-ACP) were combined to synthesize a novel nanomaterial, carboxylated polyamidoamine dendrimer/amorphous calcium phosphate nanocomposite (PAMAM-COOH/ACP). This article aims to evaluate the remineralization effect of PAMAM-COOH/ACP of dentin type I collagen fibrils in vitro. Fluorescence labeling technique was innovatively used to observe and evaluate the remineralization effect. PAMAM-COOH/ACP showed superior remineralization ability of human dentin type I collagen fibrils, especially the intrafibrillar remineralization. Therefore, the novel nanomaterial PAMAM-COOH/ACP is promising to prevent and treat various diseases caused by dentin demineralization and to improve various dental materials.

Access to Bromo-γ-butenolides via Zwitterion-Catalyzed Rearrangement of Cyclopropene Carboxylic Acids.

γ-Butenolides are useful structural motifs in many pharmaceutically relevant compounds. In particular, halogenated γ-butenolides are attractive building blocks because the halogen handles can readily be manipulated to give various functional molecules. In this study, a catalytic synthesis of halogenated γ-butenolides from cyclopropene carboxylic acids was developed using zwitterionic catalysts and N-haloamides as the halogen sources. The catalytic protocol could also be applied to the synthesis of halogenated pyrrolones by using cyclopropene amides as the starting materials.

Construction of a high-density linkage map and detection of sex-specific markers in Penaeus japonicus.

Penaeus japonicus is one of the most important farmed shrimp species in many countries. Sexual dimorphism is observed in P. japonicus, in which females grow faster and larger than males; therefore, a unisexual female culture of P. japonicus could improve the efficiency of productivity. However, the genetic mechanisms underlying sex determination in P. japonicus are unclear. In this study, we constructed a high-density genetic linkage map of P. japonicus using genotyping-by-sequencing (GBS) technology in a full-sib family. The final map was 3,481.98 cM in length and contained 29,757 single nucleotide polymorphisms (SNPs). These SNPs were distributed on 41 sex-averaged linkage groups, with an average inter-marker distance of 0.123 cM. One haplotype, harboring five sex-specific SNPs, was detected in linkage group 1 (LG1), and its corresponding confidence interval ranged from 211.840 to 212.592 cM. Therefore, this high-density genetic linkage map will be informative for genome assembly and marker-assisted breeding, and the sex-linked SNPs will be helpful for further studies on molecular mechanisms of sex determination and unisexual culture of P. japonicus in the future.

Define dose field to assess the modulation complexity of intensity-modulated radiation therapy.

PURPOSE: Introduce a new concept of dose field to assess the modulation complexity (MC) of intensity-modulated radiation therapy (IMRT). METHODS: A total of 91 IMRT plans for different diseases were retrospectively retrieved randomly from treatment database. The dose field of plans were calculated and feature values such as force magnitude and diversity were defined and extracted. Correlation analysis between these feature values and execution cost, delivery accuracy of plans was performed, to verify the validity of dose field in characterizing the MC. RESULTS: The feature values of dose field in different disease own significant differences (p < 0.001). For correlation analysis, number of control point (CP) and cumulative perimeter of CP have the highest correlation with angle entropy (0.815 and 0.848 respectively), while the correlation between number of monitor units(MU), cumulative area of CP and force, force entropy is higher than others (0.797-0.909). However, complexity of CP shape is almost irrelevant to all the dose field features. The gamma passing rate and the dose field features shows a weak negative correlation trend. CONCLUSIONS: Dose field can be used as a tool to assess the MC of IMRT.

2'3'-cGAMP triggers a STING- and NF-κB-dependent broad antiviral response in Drosophila.

We previously reported that an ortholog of STING regulates infection by picorna-like viruses in Drosophila In mammals, STING is activated by the cyclic dinucleotide 2'3'-cGAMP produced by cGAS, which acts as a receptor for cytosolic DNA. Here, we showed that injection of flies with 2'3'-cGAMP induced the expression of dSTING-regulated genes. Coinjection of 2'3'-cGAMP with a panel of RNA or DNA viruses resulted in substantially reduced viral replication. This 2'3'-cGAMP-mediated protection was still observed in flies with mutations in Atg7 and AGO2, genes that encode key components of the autophagy and small interfering RNA pathways, respectively. By contrast, this protection was abrogated in flies with mutations in the gene encoding the NF-κB transcription factor Relish. Transcriptomic analysis of 2'3'-cGAMP-injected flies revealed a complex response pattern in which genes were rapidly induced, induced after a delay, or induced in a sustained manner. Our results reveal that dSTING regulates an NF-κB-dependent antiviral program that predates the emergence of interferons in vertebrates.

Patterning Graphene Films by H2O-Based Magnetic-Assisted UV Photolysis.

Properly cutting graphene into certain high-quality micro-/nanoscale structures in a cost-effective way has a critical role. Here, we report a novel approach to pattern graphene films by H2O-based magnetic-assisted ultraviolet (UV) photolysis under irradiation at 184.9 nm. By virtue of the paramagnetic characteristic, the photo-dissociated hydroxyl [OH(X2Π)] radicals are magnetized and have their oxidation capability highly enhanced through converting into an accelerated directional motion. Meanwhile, the precursor of H2O(X̃1A1) molecules distributes uniformly thanks to its weak diamagnetic characteristic, and there exists no instable diamagnetic intermediate to cause lateral oxidation. Possessing these unique traits, the H2O-based magnetic-assisted UV photolysis has the capability of making graphene microscale patterns with the linewidth down to 8.5 µm under a copper grid shadow mask. Furthermore, it is feasible to pattern graphene films into 40 nm-wide ribbons under ZnO nanowires and realize hybrid graphene/ZnO nanoribbon field-effect transistors with a hole mobility up to 7200 cm2·V-1·s-1. The X-ray photoelectron spectroscopy and time-of-flight secondary ion mass spectrometry analyses reveal that OH(X2Π) radicals act as a strong oxidant and that another product of H(12S) adsorbs weakly on graphene.

Apolipoprotein M, identified as a novel hepatitis C virus (HCV) particle associated protein, contributes to HCV assembly and interacts with E2 protein.

Hepatitis C virus (HCV) infection is a major cause of chronic liver diseases such as steatosis, cirrhosis, and hepatocellular carcinoma. HCV particles have been found to associate with apolipoproteins, and apolipoproteins not only participate in the HCV life cycle, but also help HCV escape recognition by the host immune system, which pose challenges for the development of both HCV treatments and vaccines. However, no study has reported on the comprehensive identification of apolipoprotein associations with HCV particles. In the present study, we performed proteome analysis by affinity purification coupled with mass spectrometry (AP-MS) to comprehensively identify the apolipoprotein associations with HCV particles, and ApoM was first identified by AP-MS besides the previously reported ApoE, ApoB, ApoA-I and ApoC-I. Additionally, three assays further confirmed that ApoM was a novel virus particle associated protein. We also showed that ApoM was required for HCV production, especially for the assembly/release step of HCV life cycle. Furthermore, ApoM interacted with the HCV E2 protein. Finally, HCV infection reduced ApoM expression both in vitro and in vivo. Collectively, our study demonstrates that ApoM, identified as a novel HCV particle associated protein, contributes to HCV assembly/release and interacts with HCV E2 protein. It provides new insights on how HCV and the host apolipoproteins are reciprocally influenced and lays a basis for research in developing innovative antiviral strategies.

Eccentric magnetic microcapsules for MRI-guided local administration and pH-regulated drug release.

In this work, we present a novel class of uniform eccentric magnetic microcapsules based on polydimethylsiloxane (PDMS) for magnetic resonance imaging (MRI)-guided local injection and pH-regulated drug release. The microcapsules contained magnetic nanoparticles in their PDMS shells, which allowed them to be easily tracked by MRI during administration. Besides, they showed pH-dependent drug release profiles due to dissolution of the embedded magnetic nanoparticles in acidic solutions. Moreover, by tuning the mass fraction of the magnetic nanoparticles, we could further regulate the release rate of drug molecules from them. As a demonstration, we investigated the delivery of cis-platinum using the microcapsules through an in vitro cell test, which confirmed the pH-controlled release of the drug in phantom tissues. Our study suggests that this type of eccentric magnetic microcapsules could be simultaneously employed as a potential imaging contrast and a smart drug delivery system, holding great potential for guided local therapy.

Autologous and Nonautologous Blood Transfusion in Patients with Ruptured Ectopic Pregnancy and Severe Blood Loss.

BACKGROUND: There are some theoretical concerns for the use of intraoperative cell salvage (ICS) in patients with ectopic pregnancy. This study aimed to observe the impact of ICS on the coagulation function and clinical outcomes of patients with ruptured ectopic pregnancy and severe blood loss. METHODS: This was a retrospective study of 225 patients with ruptured ectopic pregnancy and severe blood loss treated at the Third Affiliated Hospital of Guangxi Medical University between January 2012 and May 2016. Patients were grouped according to ICS (n = 116) and controls (n = 109, allogenic transfusion and no transfusion). RESULTS: Compared with controls, patients with ICS had shorter hospitalization (P = 0.007), lower requirement for allogenic blood products (P < 0.001), and higher hemoglobin levels at discharge (P < 0.001). There were no complications/ adverse reactions. In the ICS group, hemoglobin at discharge (-6.5%, P = 0.002) and thrombin time (-3.7%, P = 0.002) were decreased 24 h after surgery, while 24 h APTT was increased (+4.6%, P < 0.001). In the control group, hemoglobin at discharge (-16.8%, P < 0.001) was decreased after surgery and 24 h APTT was increased (+2.4%, P = 0.045). At discharge, hemoglobin levels were higher in the ICS group (P < 0.001). CONCLUSION: ICS was associated with good clinical outcomes in patients with ruptured ectopic pregnancy and severe blood loss.