Dendritic boron and nitrogen doped high-entropy alloy porous carbon fibers for high-efficiency hydrogen evolution reaction.

Among various electrocatalysts, high-entropy alloys (HEAs) have gained significant attention for their unique properties and excellent catalytic activity in the hydrogen evolution reaction (HER). However, the precise synthesis of HEA catalysts in small sizes remains challenging, which limits further improvement in their catalytic performance. In this study, boron- and nitrogen-doped HEA porous carbon nanofibers (HE-BN/PCNF) with an in situ-grown dendritic structure were successfully prepared, inspired by the germination and growth of tree branches. Furthermore, the dendritic fibers constrained the growth of HEA particles, leading to the synthesis of quantum dot-sized (1.67 nm) HEA particles, which also provide a pathway for designing HEA quantum dots in the future. This work provides design ideas and guiding suggestions for the preparation of borated HEA fibers with different elemental combinations and for the application of dendritic nanofibers in various fields.

Loading cinnamon oil into pH-sensitive chitosan grafted mesoporous silica nanoparticles via supercritical carbon dioxide.

Stimulus-responsive mesoporous silica nanoparticles (MSNs) have displayed great potentiality for controlled-release and targeted drug delivery. In the current work, a supercritical fluid method was utilized to successfully prepare cinnamon oil loaded into chitosan grafted MSNs (CO@CS-MSNs). The influencing factors of drug loads, such as pressure, temperature, impregnation time and depressure time, were investigated. The structure of CO@CS-MSNs was demonstrated with Fourier-transform infrared (FT-IR) spectroscopy, transmission electron microscope (TEM), scanning electron microscopy (SEM), thermogravimetry (TG) as well as X-ray diffraction (XRD). The drug release assays in vitro at various pH conditions displayed that CO@CS-MSNs had an excellent pH-responsive release behavior, which confirmed that CO was loaded successfully into the CO@CS-MSNs. The findings indicated that the supercritical fluid approach is a non-destructive and efficient approach for stimulus-responsive MSNs, which is expected to further expand its application range.

Malformation of the endoplasmic reticulum system evolving into giant inclusions and Auer bodies in acute promyelocytic leukemia: an ultrastructural study of 6 cases.

The endoplasmic reticulum（ER）is the largest membranous network serving as a region for protein, lipid and steroid synthesis, transport and storage. Detailed information about ER-cisternae, ER-tubules and rough endoplasmic reticulum (rER) is scarce in human blood cells. This study describes a series of giant inclusions and Auer bodies in promyeloblasts in six patients with acute promyelocytic leukemia (APL), by light microscopy, transmission electron microscopy (TEM) and cytochemical stains. TEM revealed that giant inclusions and pro-Auer bodies were associated with rER and surrounded by tubular structures composed of degenerated or redundant membrane in promyeloblasts, which corresponded with elements of the ER system. This paper reveals that in the promyeloblasts of APL, ER is the source of and transforms progressively into giant inclusions and Auer bodies.

Viral RNA capping: Mechanisms and antiviral therapy.

RNA capping is an essential trigger for protein translation in eukaryotic cells. Many viruses have evolved various strategies for initiating the translation of viral genes and generating progeny virions in infected cells via synthesizing cap structure or stealing the RNA cap from nascent host messenger ribonucleotide acid (mRNA). In addition to protein translation, a new understanding of the role of the RNA cap in antiviral innate immunity has advanced the field of mRNA synthesis in vitro and therapeutic applications. Recent studies on these viral RNA capping systems have revealed startlingly diverse ways and molecular machinery. A comprehensive understanding of how viruses accomplish the RNA capping in infected cells is pivotal for designing effective broad-spectrum antiviral therapies. Here we systematically review the contemporary insights into the RNA-capping mechanisms employed by viruses causing human and animal infectious diseases, while also highlighting its impact on host antiviral innate immune response. The therapeutic applications of targeting RNA capping against viral infections and the development of RNA-capping inhibitors are also summarized.

A novel mouse model carrying a gene trap insertion into the Hmgxb4 gene locus to examine Hmgxb4 expression in vivo.

HMG (high mobility group) proteins are a diverse family of nonhistone chromosomal proteins that interact with DNA and a wide range of transcriptional regulators to regulate the structural architecture of DNA. HMGXB4 (also known as HMG2L1) is an HMG protein family member that contains a single HMG box domain. Our previous studies have demonstrated that HMGXB4 suppresses smooth muscle differentiation and exacerbates endotoxemia by promoting a systemic inflammatory response in mice. However, the expression of Hmgxb4 in vivo has not fully examined. Herein, we generated a mouse model that harbors a gene trap in the form of a lacZ gene insertion into the Hmgxb4 gene. This mouse enables the visualization of endogenous HMGXB4 expression in different tissues via staining for the ß-galactosidase activity of LacZ which is under the control of the endogenous Hmgxb4 gene promoter. We found that HMGXB4 is widely expressed in mouse tissues and is a nuclear protein. Furthermore, the Hmgxb4 gene trap mice exhibit normal cardiac function and blood pressure. Measurement of ß-galactosidase activity in the Hmgxb4 gene trap mice demonstrated that the arterial injury significantly induces Hmgxb4 expression. In summary, the Hmgxb4 gene trap reporter mouse described here provides a valuable tool to examine the expression level of endogenous Hmgxb4 in both physiological and pathological settings in vivo.

BAG6 inhibits influenza A virus replication by inducing viral polymerase subunit PB2 degradation and perturbing RdRp complex assembly.

The interaction between influenza A virus (IAV) and host proteins is an important process that greatly influences viral replication and pathogenicity. PB2 protein is a subunit of viral ribonucleoprotein (vRNP) complex playing distinct roles in viral transcription and replication. BAG6 (BCL2-associated athanogene 6) as a multifunctional host protein participates in physiological and pathological processes. Here, we identify BAG6 as a new restriction factor for IAV replication through targeting PB2. For both avian and human influenza viruses, overexpression of BAG6 reduced viral protein expression and virus titers, whereas deletion of BAG6 significantly enhanced virus replication. Moreover, BAG6-knockdown mice developed more severe clinical symptoms and higher viral loads upon IAV infection. Mechanistically, BAG6 restricted IAV transcription and replication by inhibiting the activity of viral RNA-dependent RNA polymerase (RdRp). The co-immunoprecipitation assays showed BAG6 specifically interacted with the N-terminus of PB2 and competed with PB1 for RdRp complex assembly. The ubiquitination assay indicated that BAG6 promoted PB2 ubiquitination at K189 residue and targeted PB2 for K48-linked ubiquitination degradation. The antiviral effect of BAG6 necessitated its N-terminal region containing a ubiquitin-like (UBL) domain (17-92aa) and a PB2-binding domain (124-186aa), which are synergistically responsible for viral polymerase subunit PB2 degradation and perturbing RdRp complex assembly. These findings unravel a novel antiviral mechanism via the interaction of viral PB2 and host protein BAG6 during avian or human influenza virus infection and highlight a potential application of BAG6 for antiviral drug development.

Association of maternal blood high-mobility group box 1 levels and adverse pregnancy outcomes: A systematic review and meta-analysis.

Conflicting findings have emerged regarding the levels of high mobility group box 1 (HMGB1) in individuals experiencing adverse pregnancy outcomes. Here we conducted a meta-analysis to assess the association between maternal blood HMGB1 levels and adverse pregnancy outcomes. Utilizing databases such as PubMed, Cochrane Central Register of Controlled Trials, Web of Science, Embase and China National Knowledge Infrastructure (CNKI), a systematic literature search was conducted in January 2024. Eligible literature was screened according to inclusion and exclusion criteria. Quality assessment was evaluated using the Newcastle-Ottawa Scale (NOS). The extracted data were analyzed using Review Manager 5.4 and STATA 12.0 software. 21 observational studies with a total of 2471 participants were included in this meta-analysis. Significantly higher peripheral blood levels of HMGB1 were associated with preeclampsia (PE) (SMD=1.34; 95% CI: 0.72-1.95; P < 0.0001) and gestational diabetes mellitus (GDM) (SMD=1.20; 95% CI: 0.31-2.09; P = 0.009). Additionally, HMGB1 levels in peripheral blood were significantly elevated in patients with unexplained recurrent spontaneous abortion (URSA) than those in pregnancy controls (SMD=4.22; 95% CI: 1.64-6.80; P = 0.001) or non-pregnancy controls (SMD=3.87; 95% CI: 1.81-5.92; P = 0.0002). Interestingly, higher blood HMGB1 levels were observed in women with preterm birth (PTB), however, the results did not reach a statistical difference (SMD=0.54; 95% CI: -0.36-1.44; P = 0.24). In conclusion, overexpressed maternal blood HMGB1 levels were associated with adverse pregnancy outcomes, including PE, GDM and URSA. Further studies should be conducted to validate the efficacy of HMGB1 as a biomarker for assessing the risk of adverse pregnancy outcomes.

Spatiotemporal genotype replacement of H5N8 avian influenza viruses contributed to H5N1 emergence in 2021/2022 panzootic.

Since 2020, clade 2.3.4.4b highly pathogenic avian influenza H5N8 and H5N1 viruses have swept through continents, posing serious threats to the world. Through comprehensive analyses of epidemiological, genetic, and bird migration data, we found that the dominant genotype replacement of the H5N8 viruses in 2020 contributed to the H5N1 outbreak in the 2021/2022 wave. The 2020 outbreak of the H5N8 G1 genotype instead of the G0 genotype produced reassortment opportunities and led to the emergence of a new H5N1 virus with G1's HA and MP genes. Despite extensive reassortments in the 2021/2022 wave, the H5N1 virus retained the HA and MP genes, causing a significant outbreak in Europe and North America. Furtherly, through the wild bird migration flyways investigation, we found that the temporal-spatial coincidence between the outbreak of the H5N8 G1 virus and the bird autumn migration may have expanded the H5 viral spread, which may be one of the main drivers of the emergence of the 2020-2022 H5 panzootic.IMPORTANCESince 2020, highly pathogenic avian influenza (HPAI) H5 subtype variants of clade 2.3.4.4b have spread across continents, posing unprecedented threats globally. However, the factors promoting the genesis and spread of H5 HPAI viruses remain unclear. Here, we found that the spatiotemporal genotype replacement of H5N8 HPAI viruses contributed to the emergence of the H5N1 variant that caused the 2021/2022 panzootic, and the viral evolution in poultry of Egypt and surrounding area and autumn bird migration from the Russia-Kazakhstan region to Europe are important drivers of the emergence of the 2020-2022 H5 panzootic. These findings provide important targets for early warning and could help control the current and future HPAI epidemics.

Recombinant parainfluenza virus 5 expressing clade 2.3.4.4b H5 hemagglutinin protein confers broad protection against H5Ny influenza viruses.

The global circulation of clade 2.3.4.4b H5Ny highly pathogenic avian influenza viruses (HPAIVs) in poultry and wild birds, increasing mammal infections, continues to pose a public health threat and may even form a pandemic. An efficacious vaccine against H5Ny HPAIVs is crucial for emergency use and pandemic preparedness. In this study, we developed a parainfluenza virus 5 (PIV5)-based vaccine candidate expressing hemagglutinin (HA) protein of clade 2.3.4.4b H5 HPAIV, termed rPIV5-H5, and evaluated its safety and efficacy in mice and ferrets. Our results demonstrated that intranasal immunization with a single dose of rPIV5-H5 could stimulate H5-specific antibody responses, moreover, a prime-boost regimen using rPIV5-H5 stimulated robust humoral, cellular, and mucosal immune responses in mice. Challenge study showed that rPIV5-H5 prime-boost regimen provided sterile immunity against lethal clade 2.3.4.4b H5N1 virus infection in mice and ferrets. Notably, rPIV5-H5 prime-boost regimen provided protection in mice against challenge with lethal doses of heterologous clades 2.2, 2.3.2, and 2.3.4 H5N1, and clade 2.3.4.4h H5N6 viruses. These results revealed that rPIV5-H5 can elicit protective immunity against a diverse clade of highly pathogenic H5Ny virus infection in mammals, highlighting the potential of rPIV5-H5 as a pan-H5 influenza vaccine candidate for emergency use.IMPORTANCEClade 2.3.4.4b H5Ny highly pathogenic avian influenza viruses (HPAIVs) have been widely circulating in wild birds and domestic poultry all over the world, leading to infections in mammals, including humans. Here, we developed a recombinant PIV5-vectored vaccine candidate expressing the HA protein of clade 2.3.4.4b H5 virus. Intranasal immunization with rPIV5-H5 in mice induced airway mucosal IgA responses, high levels of antibodies, and robust T-cell responses. Importantly, rPIV5-H5 conferred complete protection in mice and ferrets against clade 2.3.4.4b H5N1 virus challenge, the protective immunity was extended against heterologous H5Ny viruses. Taken together, our data demonstrate that rPIV5-H5 is a promising vaccine candidate against diverse H5Ny influenza viruses in mammals.

Isolation and characterization of genotype 4 Eurasian avian-like H1N1 influenza virus in pigs suffering from pneumonia.

Swine influenza viruses pose ongoing threat to pork industry throughout the world. In 2023, fattening pigs from a swine farm in Inner Mongolia of China experienced influenza-like symptoms. Co-infection of influenza A virus with Pasteurella multocida was diagnosed in lung tissues of diseased pigs and a genotype 4 (G4) Eurasian avian-like (EA) H1N1 virus was isolated, which was named as A/swine/Neimenggu/0326/2023. We demonstrated the virus preferentially bound human-like SAα2,6Gal receptor. It was noteworthy that the virus possessed multiple genetic markers for mammalian adaptation in the internal genes. Animal studies showed that compared with genotype 1 (G1) EA H1N1 virus and early prevalent G4 EA H1N1 virus, A/swine/Neimenggu/0326/2023 virus exhibited increased virus shedding, enhanced replication in lungs, and caused more severe lung lesions in pigs. These findings indicate that the G4 EA H1N1 virus poses increased threat to pork industry, controlling the prevailing viruses in pigs should be promptly implemented.

A new chromosome-scale duck genome shows a major histocompatibility complex with several expanded multigene families.

BACKGROUND: The duck (Anas platyrhynchos) is one of the principal natural hosts of influenza A virus (IAV), harbors almost all subtypes of IAVs and resists to many IAVs which cause extreme virulence in chicken and human. However, the response of duck's adaptive immune system to IAV infection is poorly characterized due to lack of a detailed gene map of the major histocompatibility complex (MHC). RESULTS: We herein reported a chromosome-scale Beijing duck assembly by integrating Nanopore, Bionano, and Hi-C data. This new reference genome SKLA1.0 covers 40 chromosomes, improves the contig N50 of the previous duck assembly with highest contiguity (ZJU1.0) of more than a 5.79-fold, surpasses the chicken and zebra finch references in sequence contiguity and contains a complete genomic map of the MHC. Our 3D MHC genomic map demonstrated that gene family arrangement in this region was primordial; however, families such as AnplMHCI, AnplMHCIIß, AnplDMB, NKRL (NK cell receptor-like genes) and BTN underwent gene expansion events making this area complex. These gene families are distributed in two TADs and genes sharing the same TAD may work in a co-regulated model. CONCLUSIONS: These observations supported the hypothesis that duck's adaptive immunity had been optimized with expanded and diversified key immune genes which might help duck to combat influenza virus. This work provided a high-quality Beijing duck genome for biological research and shed light on new strategies for AIV control.

Simultaneous preconcentration and quantification of ultra-trace tin and lead species in seawater by online SPE coupled with HPLC-ICP-MS.

BACKGROUND: Tin and lead contamination is a global threat to marine ecosystems considering their species-specific toxicity, bioavailability and mobility. Hence simultaneous measurement of multiple tin and lead compounds at µg L-1 to pg L-1 levels in environmental water is always an indispensable but challengeable task. High performance liquid chromatography coupled with inductively coupled plasma mass spectrometry (HPLC-ICP-MS) is one of the most widely used choices for this purpose because of good sensitivity, strong separation power and good compatibility. Previous HPLC-ICP-MS methods based on a single elemental speciation strategy are low-efficiency and sensitivity-insufficient for a large set of unstable samples and interaction of multiple metal(loid)s down to ng L-1 levels. RESULTS: In this study, we developed a sensitive, efficient and environment-friendly analytical method for accurate quantification of inorganic and organic species of tin and lead simultaneously based on HPLC-ICP-MS with online integration of solid phase extraction (SPE). By using graphene oxide modified silica conditioned with 1 mM benzoic acid to enrich tin and lead species from 10 mL sample, detection limits were improved to 2-8 pg per liter due to satisfactory enrichment factors (522-2848 folds). The SPE-HPLC-ICP-MS method was applicable to quantification of ultra-trace tin and lead species at pg L-1 levels in uncontaminated seawater. Tributyltin was the only tin species detected at subnanograms per liter levels while Pb(II) was the only lead species detected at several nanograms per liter in thirteen coastal seawater samples collected in Hangzhou Bay, indicating light contamination of tin and lead. SIGNIFICANCE: Overall, the proposed SPE-HPLC-ICP-MS method is highly sensitive, efficient and environment-friendly that are fairly suitable to routine speciation analysis of tin and lead in environmental, food, and biological samples.

Simultaneous quantification of tin and lead species in Antarctic krill and fish by interfacing high-performance liquid chromatography with inductively coupled plasma mass spectrometry based on strong cation-exchange and Amphion columns.

Tin and lead are a global concern considering their species-dependent toxicity, bioavailability and transformation. Simultaneous speciation analysis of tin and lead is challenging for a large food capacity containing unstable species. Herein, we developed two sensitive methods for rapid quantification of tin and lead species in Antarctic seafood by high-performance liquid chromatography and inductively coupled plasma mass spectrometry based on strong cation-exchange and Amphion columns. Inorganic tin and lead, four organotin and two organolead compounds can be analysed in 16 min on a 10-cm Amphion II column (mobile phase: 4 mM sodium dodecyl benzene sulfonate at pH 2.0) with 0.02-0.24 µg L-1 detection limits. The method was applied to Antarctic krill and fish, demonstrating the presence of any tin and lead species down to µg kg-1 level. Overall, the proposed methods are sensitive, efficient and environment-friendly for routine speciation analysis of tin and lead in food samples.

Plasmon enhanced luminescence of Tb/Eu co-doped film by Au NRs-PVA nanocomposite film.

Plasmonic nanostructures have great potential for improving the radiation properties of emitters. Here, the plasmonic Au nanorods-PVA nanocomposite films are used to uniformly improve the photoluminescence of Tb/Eu co-doped PMMA film within the local micro-region. Under the excitation of 292 nm, the maximum enhancement factor is 37.2-fold for emission at 612 nm and 21.6-fold for emission at 545 nm. Moreover, the finite different time domain simulations are developed to further explain the experimental results. It is indicated that the modulation of luminescence can be attributed to the increase of the local density of optical states through the Purcell effect and the improvement of the energy transfer efficiency between Tb and Eu. Under the excitation of 360 nm, the maximum enhancement factor is about 71.5-fold. In this case, the Au nanorods are mainly used for modulating the emission process at 612 nm, which deduced a greater enhancement factor at 612 nm. This study provides a deep understanding of the interactions between rare earth ions co-doped materials and plasmonic nanostructures, building a bridge to fabricate a useful platform for several applications, such as thin film-based detectors and sensors.

Multiscale spatial-temporal evolution of energy carbon footprint in the Yellow River Basin of China based on DMSP/OLS and NPP/VIIRS integrated data.

Increased CO2 emissions from urban energy consumption pose a significant challenge to regional carbon mitigation policies. In this paper, we integrated two nighttime light (NTL) data: the Defense Meteorological Satellite Program (DMSP) Operational Linescan System (OLS) and the Visible Infrared Imaging Radiometer Suite (VIIRS) on the Suomi National Polar-orbiting Partnership (NPP) composite data to estimate the energy carbon emissions from 2000 to 2019. Then the spatiotemporal dynamics of carbon footprint and deficit in the Yellow River Basin were analyzed at the provincial, municipal, and county scales combined with NPP data. The study shows that (1) the total amount of energy consumption CO2 emissions in the Yellow River Basin had increased from 1332 Mt in 2000 to 6469 Mt in 2019, but the average annual growth rate slowed down after 2010 from 11.5 to 5.61%. (2) From 2000 to 2018, the provinces with the highest carbon footprint and carbon deficit were concentrated in Inner Mongolia and Shanxi. In 2018, Inner Mongolia's carbon footprint was 1366.91 × 104 km2, accounting for 22.8% of the total. Cities with high carbon footprint were mainly economic centers and energy-intensive areas of various provinces. High-carbon deficit counties were mainly distributed in the western region. In 2018, 954 counties exhibited carbon deficits. (3) The carbon footprint in the Yellow River Basin at the municipal and county scales have a significant spatial correlation. The H-H clusters of the carbon footprint on the municipal scale were distributed in the middle reaches of the Yellow River Basin. At the county scale, the L-L clusters were mainly in Sichuan and eastern Henan regions. Through the analysis of the spatial and temporal evolution of carbon footprint and carbon deficit in the Yellow River Basin, it is significant to measure the degree of comprehensive coordination of carbon sources and sinks in the basin, to grasp the differences in the level of regional carbon emissions, and to promote synergistic regional governance, assist in the formulation of more precise carbon emission reduction policies, and to promote green and high-quality development.

Semi-Supervised Medical Image Segmentation Using Cross-Style Consistency With Shape-Aware and Local Context Constraints.

Despite the remarkable progress in semi-supervised medical image segmentation methods based on deep learning, their application to real-life clinical scenarios still faces considerable challenges. For example, insufficient labeled data often makes it difficult for networks to capture the complexity and variability of the anatomical regions to be segmented. To address these problems, we design a new semi-supervised segmentation framework that aspires to produce anatomically plausible predictions. Our framework comprises two parallel networks: shape-agnostic and shape-aware networks. These networks learn from each other, enabling effective utilization of unlabeled data. Our shape-aware network implicitly introduces shape guidance to capture shape fine-grained information. Meanwhile, shape-agnostic networks employ uncertainty estimation to further obtain reliable pseudo-labels for the counterpart. We also employ a cross-style consistency strategy to enhance the network's utilization of unlabeled data. It enriches the dataset to prevent overfitting and further eases the coupling of the two networks that learn from each other. Our proposed architecture also incorporates a novel loss term that facilitates the learning of the local context of segmentation by the network, thereby enhancing the overall accuracy of prediction. Experiments on three different datasets of medical images show that our method outperforms many excellent semi-supervised segmentation methods and outperforms them in perceiving shape. The code can be seen at https://github.com/igip-liu/SLC-Net.

Structural characterization and origin of surface vesicles in monocytes: another membranous pathway from cytoplasm to cell surface.

The monocytes in acute monocytic leukemia (AML-M5b) were analyzed by scanning and transmission electron microscopy (SEM and TEM) to understand more fully their structure and origin. By SEM, monocytes exhibited localized expansions of the surface, some of which appeared to bud off as surface vesicles (SVs). Filopodial processes and pseudopodia were also present. TEM demonstrated that the SVs were composed of a double-membrane at the pole away from the cell body, and a single membrane nearer to the cell body. In the peripheral cytoplasm, intracellular vesicles (IVs) had the appearance of vacuoles and were enclosed by single membranes. Most SVs were characterized by a notch as a rER edge and an expanded head. Filopodial processes had the same thickness of 40 nm as the SV walls, which suggested a close developmental relationship between the two. Pseudopodia between SVs were irregular in size. Rod-like rER cisternae were prominent in the peripheral cytoplasm and some showed a close physical juxtaposition as to suggest a transition from rER to IVs to SVs. Ultrastructural cytochemistry demonstrated activity of 5'-nucleotidase over rER, SVs, filopodial processes and pseudopodia, and a patchy reaction over other areas of plasma membrane. Overall, the results indicated that rER transforms into SVs, filopodial processes and pseudopodia, as a way of integrating cytoplasmic membranes into the plasma membrane.

An inhibitor-monitorable single-tube duplex quantitative real-time PCR assay for the detection of Candidatus Liberibacter asiaticus.

Huanglongbing (HLB) is a citrus infectious disease caused by Candidatus Liberibacters spp. Recently, it has begun to spread rapidly worldwide, causing significant losses to the citrus industry. Early diagnosis of HLB relies on quantitative real-time PCR assays. However, the PCR inhibitors found in the nucleic acid extracted from plant materials pose challenges for PCR assays because they may result in false-negative results. Internal standard (IS) can be introduced to establish a single-tube duplex PCR (STD-PCR) for monitoring the influence of the PCR-inhibitor, but it also brings the risk of false-negative results because the amplification of IS may compete with the target. To solve this problem, we proposed a mutation-enhanced single-tube duplex PCR (mSTD-PCR) containing IS with mutant-type primers. By introducing the 3'-terminal mutation in the primer of IS to weaken its amplification reaction and its inhibition of Candidatus Liberibacter asiaticus (CLas) detection, the sensitivity and quantitative accuracy of CLas detection will not be affected by IS. In evaluating the sensitivity of CLas detection using simulation samples, the mSTD-PCR showed consistent sensitivity at 25 copies/test compared with the single-plex CLas assay. The detection result of 30 leaves and 30 root samples showed that mSTD-PCR could recognize false-negative results caused by the PCR inhibitors and reduce workload by 48% compared with the single-plex CLas assay. Generally, the proposed mSTD-PCR provides a reliable, efficient, inhibitor-monitorable, and quantitative screening method for accurately controlling HLB and a universal method for establishing PCR assay for various pathogens.

Is Reaction Acceleration of Microdroplet Chemistry Favorable to Controlling the Enantioselectivity?

Microdroplet chemistry has been proven to amazingly accelerate many chemical and biological reactions in the past 2 decades. Current microdroplet accelerated reactions are predominantly symmetric synthetic but minorly asymmetric synthetic reactions, where stereoselectivity is scarcely concerned. This study selected unimolecular and bimolecular reactions, multicomponent Passerini reactions, and enzymatic ketone reduction as the model reactions to illustrate whether reaction acceleration of microdroplet chemistry is favorable to retaining a chiral center and controlling the enantioselectivity or not. The results illustrated that microdroplet chemistry did not disrupt pre-existing stereogenic centers in chiral starting materials during reactions but did harm to stereospecificity in asymmetric catalysis by chiral catalysts and chiral organic ligands with the exclusion of enzymatic reactions. Our preliminary study reminds us of more cautions to the product enantioselectivity when conducting asymmetric catalysis in microdroplets. We also hope this study may promote more valuable further research on the stereoselectivity of microdroplet chemistry.