Zipper-Confined DNA Nanoframe for High-Efficient and High-Contrast Imaging of Heterogeneous Tumor Cell.

Current study in the heterogeneity and physiological behavior of tumor cells is limited by the fluorescence in situ hybridization technology in terms of probe assembly efficiency, background suppression capability, and target compatibility. In a typically well-designed assay, hybridization probes are constructed in a confined nanostructure to achieve a rapid assembly for efficient signal response, while the excessively high local concentration between different probes inevitably leads to nonspecific background leakage. Inspired by the fabric zipper, we propose a novel confinement reaction pattern in a zipper-confined DNA nanoframe (ZCDN), where two kinds of hairpin probes are independently anchored respective tracks. The metastable states of the dual tracks can well avoid signal leakage caused by the nonspecific probe configuration change. Biomarker-mediated proximity ligation reduces the local distance of dual tracks, kinetically triggering an efficient allosteric chain reaction between the hairpin probes. This method circumvents nonspecific background leakage while maintaining a high efficiency in responding to targets. ZCDN is employed to track different cancer biomarkers located in both the cytoplasm and cytomembrane, of which the expression level and oligomerization behavior can provide crucial information regarding intratumoral heterogeneity. ZCDN exhibits high target response efficiency and strong background suppression capabilities and is compatible with various types of biological targets, thus providing a desirable tool for advanced molecular diagnostics.

Inhibition of histone acetyltransferase GCN5 by a transcription factor FgPacC controls fungal adaption to host-derived iron stress.

Poaceae plants can locally accumulate iron to suppress pathogen infection. It remains unknown how pathogens overcome host-derived iron stress during their successful infections. Here, we report that Fusarium graminearum (Fg), a destructive fungal pathogen of cereal crops, is challenged by host-derived high-iron stress. Fg infection induces host alkalinization, and the pH-dependent transcription factor FgPacC undergoes a proteolytic cleavage into the functional isoform named FgPacC30 under alkaline host environment. Subsequently FgPacC30 binds to a GCCAR(R = A/G)G element at the promoters of the genes involved in iron uptake and inhibits their expression, leading to adaption of Fg to high-iron stress. Mechanistically, FgPacC30 binds to FgGcn5 protein, a catalytic subunit of Spt-Ada-Gcn5 Acetyltransferase (SAGA) complex, leading to deregulation of histone acetylation at H3K18 and H2BK11, and repression of iron uptake genes. Moreover, we identified a protein kinase FgHal4, which is highly induced by extracellular high-iron stress and protects FgPacC30 against 26S proteasome-dependent degradation by promoting FgPacC30 phosphorylation at Ser2. Collectively, this study uncovers a novel inhibitory mechanism of the SAGA complex by a transcription factor that enables a fungal pathogen to adapt to dynamic microenvironments during infection.

Seasonal ammonia emissions from an intensive beef cattle feedlot in Victoria Australia.

Ammonia (NH3) emitted from concentrated animal feeding operations can cause environmental and health problems, and indirectly contribute to greenhouse gas emissions. Cattle feedlots are known to be large sources of NH3, but few studies have documented seasonal emissions from Australian feedlots. We conducted two field campaigns to measure NH3 emissions from an intensive beef cattle feedlot in southeast Australia, and these results were combined with previous measurements at the same feedlot to document seasonal variations in emissions and to derive annual feedlot emission factors (EFs). Emission rates were calculated with an inverse dispersion modelling (IDM) technique, based on NH3 concentrations measured at the feedlot with open-path lasers (OPLs). The average area emission rates in spring, summer, autumn and winter were 90.5, 167.4, 96.2 and 86.8 µg NH3 m-2 s-1 from the cattle pens, and 22.5, 18.1, 7.7 and 20.7 µg NH3 m-2 s-1 from the manure stockpile area, respectively. The total per-animal EFs ranged from 126.0 (autumn) to 190.2 g NH3 animal-1 d-1 (summer), representing a loss of 47.5-64.6% of the fed N. Seasonal variations in emissions were related to air temperature. Slight changes in crude protein content of the cattle diet may also have impacted seasonal variability. Taking seasonal variations into consideration, the average feedlot EF was 160.4 g NH3 animal-1 d-1, with 90% of the emissions coming from the cattle pens. Extrapolating the EF to all feedlot cattle in the country, the direct NH3 emissions from Australian feedlots amount to 65.2 Gg NH3 annually, or 3.7% of the national total. Our study benchmarks seasonal and annual EFs and N losses for Australian commercial feedlots, and provides a baseline for extrapolating the impacts of mitigation efforts.

Preputial Shortening Reconstruction Surgery in a Dog with a Micropenis and Prepuce-to-Penis Size Disparity.

A 1 yr old castrated male shih tzu was referred for recurrent urinary tract infections (UTI), prostatitis, and urine dribbling that was not responsive to medical management. Physical examination and computed tomography scan revealed a micropenis with a disproportionately high prepuce-to-penis ratio. Preputial shortening with a hexagonal, full thickness preputial resection followed by preputial anastomosis was performed. The dog recovered from surgery with no complications. Urine dribbling persisted in the short-term postoperative period, but the patient achieved significant clinical improvement and resolution of his urine dribbling and recurrent UTIs at the 1 yr follow-up. In conclusion, this surgical technique was able to successfully restore quality of life in a dog with a micropenis, and preputial shortening should be considered in cases of recurrent UTIs where there is significant disparity between the size of the penis and the prepuce.

A Comparison of Laparoscopy and Laparotomy for the Management of Abdominal Trauma: A Systematic Review and Meta-analysis.

BACKGROUND: This study aimed to systematically review and compare the perioperative outcomes of laparoscopy with laparotomy for abdominal trauma patients. METHODS: We conducted a systematic review and meta-analysis comparing the perioperative outcomes of laparoscopy with laparotomy for abdominal trauma patients. Clinical endpoints included length of hospital stay, operation time, amount of intraoperative blood loss, time to postoperative exhaust, time to regular diet, time to out of bed, duration of postoperative pain, postoperative complications, perioperative mortality rate, length of intensive care unit (ICU) stay, missed injuries, conversions to laparotomy, and cure rate. RESULTS: Sixty-four studies including 9058 patients with abdominal trauma were included. In these studies, laparoscopy was used as a screening, diagnostic, or therapeutic tool. Meta-analysis showed significant reductions in the incidence of postoperative complications (relative risk [RR] [95 % confidence interval (CI)] 0.37 [0.29-0.46]), perioperative mortality rate (RR 0.64; 95 % CI 0.52-0.80), operation time (mean difference [MD] [95 % CI] -19.93 min [-34.43 to 5.43]), length of hospital stay (MD -5.15 days; 95 % CI -6.80 to 3.50), amount of intraoperative blood loss (MD -141.33 ml; 95 % CI -260.99 to 21.67), time to postoperative exhaust (MD -5.32 h; 95 % CI -8.60 to 2.05), time to regular diet (MD -3.46 h; 95 % CI -6.31 to 0.61), time to out of bed (MD -23.51 h; 95 % CI -24.85 to 22.16), duration of postoperative pain (MD -21.34 h; 95 % CI -22.65 to 20.03), length of ICU stay (MD -1.89 days; 95 % CI -4.05 to 0.27) in patients with abdominal trauma treated with laparoscopy compared with laparotomy. The pooled incidence of postoperative complications, missed injuries, conversions, and perioperative mortality rate of laparoscopy among the case reports were 0.04 (95 % CI 0.03-0.06), 0.01 (95 % CI 0.01-0.02), 0.24 (95 % CI 0.20-0.28), 0.01(95 % CI 0.01-0.02), respectively. Cure rate of laparoscopy ranged from 46 to 95 % and the pooled rate was 0.76 (95 % CI 0.71-0.81). CONCLUSIONS: Laparoscopy is an effective way to improve perioperative outcomes and reduce the complications of hemodynamically stable patients with abdominal trauma. It is worth further popularization in clinical practice.

Multimodal integration for Barrett's esophagus.

The esophageal adenocarcinoma is facing a worldwide challenge: early prediction and risk assessment in clinical Barrett's esophagus (BE). In recent years, the growing interests have been witnessed in prediction and risk assessment in clinical BE. However, the resolution is limited, and the system is huge and expensive for the existing devices. Inspired by the principle of collaboration between human eye vision and brain cortex in data processing, here we propose multimodal learning framework to tackle tasks from various modalities, which can benefit from each other. To our findings, the experimental result indicates that low-level modality can directly affect high-level modality and form the final risk grading based on contribution, which maximizes the clinical performance of medical professionals based on our findings.

The environmental and socioeconomic benefits of optimized fertilization for greenhouse vegetables.

China produces more than half of global vegetables with greenhouse farms contributes approximately 35 % to the country's overall vegetable supply. The average nitrogen (N) application rate of greenhouse vegetable production exceeds 2000 kg N ha-1 yr-1, considerably contributing to global agricultural GHG emissions and reactive N (Nr) losses. Optimizing the N fertilizer utilization in greenhouse vegetable production is essential for mitigating environmental pollution and promoting sustainable development nationally and globally. In this study, we estimated the N footprint (NF), social costs (SC, which includes ecosystem and human health damage costs caused by Nr losses to the environment) and net ecosystem economic income (NEEI, which balances between the fertilizers input cost, yield profit, and social costs) of different greenhouse vegetables (tomato, pakchoi, lettuce, cabbage) under farmers' practice (FP) and reduced fertilization treatment (R). Results showed that compared with FP, the NF of tomato, pakchoi, lettuce and cabbage in the R treatment decreased by 61 %, 29 %, 46 % and 36 %, respectively, and the social costs were decreased by 60 %, 48 %, 57 % and 50 %, respectively. On the regional scale, the reduction in N fertilizer use for greenhouse vegetables in Beijing only could save the fertilizer input cost by 1-5 million USD, and avoided SC would increase by 1-14 million USD. As a result, this increased the NEEI by 2-19million USD. This study has demonstrated that adopting reduced fertilization practices represents a cost-effective measure that not only ensures yields but also decrease social costs, NF, and improve the benefits to help achieve sustainable development of greenhouse vegetable production.

Exploration of new ways for CRISPR/Cas12a activation: DNA hairpins without PAM and toehold and single strands containing DNA and RNA bases.

The CRISPR/Cas12a system is emerging as a promising candidate for next-generation diagnostic biosensing platforms, with the discovery of new activation modes greatly expanding its applications. Here, we have identified two novel CRISPR/Cas12a system activation modes: PAM- and toehold-free DNA hairpins, and DNA-RNA hybrid strands. Utilizing a well-established real-time fluorescence method, we have demonstrated a strong correlation between DNA hairpin structures and Cas12a activation. Compared with previously reported activation modes involving single-stranded DNA and PAM-contained double-stranded DNA, the DNA hairpin activation way exhibits similar specificity and generality. Moreover, our findings indicate that increasing the number of RNA bases in DNA-RNA hybrid strands can decelerate the kinetics of Cas12a-triggered trans-cleavage of reporter probes. These newly discovered CRISPR/Cas12a activation ways hold significant potential for the development of high-performance biosensing strategies.

Split activator of CRISPR/Cas12a for direct and sensitive detection of microRNA.

CRISPR/Cas12a-based nucleic acid assays have been increasingly used for molecular diagnostics. However, most current CRISPR/Cas12a-based RNA assays require the conversion of RNA into DNA by preamplification strategies, which increases the complexity of detection. Here, we found certain chimeric DNA-RNA hybrid single strands could activate the trans-cleavage activity of Cas12a, and then discovered the activating effect of split ssDNA and RNA when they are present simultaneously. As proof of concept, split nucleic acid-activated Cas12a (SNA-Cas12a) strategy was developed for direct detection of miR-155. By adding a short ssDNA to the proximal end of the crRNA spacer sequence, we realized the direct detection of RNA targets using Cas12a. With the assistance of ssDNA, we extended the limitation that CRISPR/Cas12a cannot be activated by RNA targets. In addition, by taking advantage of the programmability of crRNA, the length of its binding to DNA and RNA was optimized to achieve the optimal efficiency in activating Cas12a. The SNA-Cas12a method enabled sensitive miR-155 detection at pM level. This method was simple, rapid, and specific. Thus, we proposed a new Cas12a-based RNA detection strategy that expanded the application of CRISPR/Cas12a.

Smart Implantable Hydrogel for Large Segmental Bone Regeneration.

Large segmental bone defects often lead to nonunion and dysfunction, posing a significant challenge for clinicians. Inspired by the intrinsic bone defect repair logic of "vascularization and then osteogenesis", this study originally reports a smart implantable hydrogel (PDS-DC) with high mechanical properties, controllable scaffold degradation, and timing drug release that can proactively match different bone healing cycles to efficiently promote bone regeneration. The main scaffold of PDS-DC consists of polyacrylamide, polydopamine, and silk fibroin, which endows it with superior interfacial adhesion, structural toughness, and mechanical stiffness. In particular, the adjustment of scaffold cross-linking agent mixing ratio can effectively regulate the in vivo degradation rate of PDS-DC and intelligently satisfy the requirements of different bone defect healing cycles. Ultimately, PDS hydrogel loaded with free desferrioxamine (DFO) and CaCO3 mineralized ZIF-90 loaded bone morphogenetic protein-2 (BMP-2) to stimulate efficient angiogenesis and osteogenesis. Notably, DFO is released rapidly by free diffusion, whereas BMP-2 is released slowly by pH-dependent layer-by-layer disintegration, resulting in a significant difference in release time, thus matching the intrinsic logic of bone defect repair. In vivo and in vitro results confirm that PDS-DC can effectively realize high-quality bone generation and intelligently regulate to adapt to different demands of bone defects.

Multiple RNA Rapid In Situ Imaging Based on Cas9 Code Key System.

Existing RNA in situ imaging strategies mostly utilize parallel repetitive nucleic acid self-assembly to achieve multiple analysis, with limitations of complicated systems and cumbersome steps. Here, a Cas9 code key system with key probe (KP) encoder and CRISPR/Cas9 signal exporter is developed. This system triggers T-protospacer adjacent motif (T-PAM structural transitions of multiple KP encoders to form coding products with uniform single-guide RNA (sgRNA) target sequences as tandem nodes. Only single sgRNA/Cas9 complex is required to cleave multiple coding products, enabling efficient "many-to-one" tandem signaling, and non-collateral cleavage activity-dependent automatic signaling output through active introduction of mismatched bases. Compared with conventional parallel multiple signaling analysis model, the proposed system greatly simplifies reaction process and enhances detection efficiency. Further, a rapid multiple RNA in situ imaging system is developed by combining the Cas9 code key system with a T-strand displacement amplification (T-SDA) signal amplifier. The constructed system is applied to tumor cells and clinicopathology slices, generating clear multi-mRNA imaging profiles in less than an hour with just one step. Therefore, this work provides reliable technical support for clinical tumor typing and molecular mechanism investigation.

mROS­calcium feedback loop promotes lethal ventricular arrhythmias and sudden cardiac death in early myocardial ischemia.

Lethal ventricular arrhythmia­sudden cardiac death (LVA­SCD) occurs frequently during the early stage of myocardial ischemia (MI). However, the mechanism underlying higher LVA­SCD incidence is still poorly understood. The present study aimed to explore the role of mitochondrial reactive oxygen species (mROS) and Ca2+ crosstalk in promoting LVA­SCD in early MI. RyR2 S2814A mice and their wild­type littermates were used. MitoTEMPO was applied to scavenge mitochondrial ROS (mROS). Mice were subjected to severe MI and the occurrence of LVA­SCD was evaluated. Levels of mitochondrial ROS and calcium (mitoCa2+), cytosolic ROS (cytoROS), and calcium (cytoCa2+), RyR2 Ser­2814 phosphorylation, CaMKII Met­282 oxidation, mitochondrial membrane potential (MMP), and glutathione/oxidized glutathione (GSH/GSSG) ratio in the myocardia were detected. Dynamic changes in mROS after hypoxia were investigated using H9c2 cells. Moreover, the myocardial phosphoproteome was analyzed to explore the related mechanisms facilitating mROS­Ca2+ crosstalk and LVA­SCD. There was a high incidence (~33.9%) of LVA­SCD in early MI. Mice who underwent SCD displayed notably elevated levels of myocardial ROS and mROS, and the latter was validated in H9c2 cells. These mice also demonstrated overloads of cytoplasmic and mitochondrial Ca2+, decreased MMP and reduced GSH/GSSG ratio, upregulated RyR2­S2814 phosphorylation and CaMKII­M282 oxidation and transient hyperphosphorylation of mitochondrial proteomes in the myocardium. mROS­specific scavenging by a mitochondria­targeted antioxidant agent (MitoTEMPO) corrected these SCD­induced alterations. S2814A mice with a genetically inactivated CaMKII phosphorylation site in RyR2 exhibited decreased overloads in cytoplasmic and mitochondrial Ca2+ and demonstrated similar effects as MitoTEMPO to correct SCD­induced changes and prevent SCD post­MI. The data confirmed crosstalk between mROS and Ca2+ in promoting LVA­SCD. Therefore, we provided evidence that there is a higher incidence of LVA­SCD in early MI, which may be attributed to a positive feedback loop between mROS and Ca2+ imbalance.

Photoredox-Catalyzed Three-Component Amidoheteroarylation of Unactivated Alkenes.

A photoredox-catalyzed 1,2-amidoheteroarylation of unactivated alkenes with O-acyl hydroxylamine derivatives and heterocycles is presented. A range of heterocycles, including quinoxaline-2(1H)-ones, azauracils, chromones, and quinolones, are capable for this process, allowing the direct synthesis of valuable heteroarylethylamine derivatives. Structurally diverse reaction substrates, including drug-based scaffolds, were successfully applied, demonstrating the practicality of this method.

Photoinduced Radical Sulfinylation of C(sp3)-H Bonds with Sulfinyl Sulfones.

A direct C(sp3)-H sulfinylation reaction of alkanes with sulfinyl sulfones via decatungstate photocatalysis is reported. The sulfinyl sulfones generated in situ from sulfinates in the presence of an acylating reagent were able to trap the alkyl radicals that were produced via the photoinduced direct hydrogen atom transfer of alkanes, leading to a range of sulfoxides. This radical sulfinylation process provides an efficient and concise method for the synthesis of sulfoxides from abundant alkanes under mild conditions. Using the same strategy, aldehydes can also be transferred to the corresponding sulfoxides via decarbonylative sulfinylation.

Effects of Psychological Distress and Coping Resources on Internet Gaming Disorder: Comparison between Chinese and Japanese University Students.

The high prevalence of Internet gaming disorder (IGD) among Asian youth indicates an urgent need to identify protective factors and examine their consistency across Asian cultures in order to facilitate cost-effective interventions. Based on the transactional theory of stress and coping, this study collected data of 1243 online gamers (45% males; 18-25 years) through an anonymous survey from universities in China and Japan and investigated whether three coping resources (i.e., mindfulness, coping flexibility, and social support) serve to protect Chinese and Japanese youth from the impact of psychological distress on IGD tendency. After adjusting for the measurement non-invariance across samples, we found that Japanese students reported higher levels of IGD tendency and psychological distress than Chinese students. The results of multiple-group SEM analyses showed that, after controlling for other predictors, mindfulness served as the strongest protective factor against IGD across samples. Moreover, the buffering effect of mindfulness on the association between psychological distress and IGD tendency of female (but not male) students was observed. Our findings highlighted the cross-cultural invariance of the impact of psychological distress and coping resources on IGD in Chinese and Japanese youth, which can be considered in future IGD prevention programs.

The autophagy-related proteins FvAtg4 and FvAtg8 are involved in virulence and fumonisin biosynthesis in Fusarium verticillioides.

Autophagy is the main intracellular degradation system by which cytoplasmic materials are transported to and degraded in the vacuole/lysosome of eukaryotic cells, and it also controls cellular differentiation and virulence in a variety of filamentous fungi. However, the contribution of the autophagic pathway to fungal development and pathogenicity in the important maize pathogen and mycotoxigenic fungus Fusarium verticillioides is still unknown. In this study, we characterized two autophagy-related proteins, FvAtg4 and FvAtg8. The F. verticillioides deletion mutants ΔFvAtg4 and ΔFvAtg8 were impaired in autophagosome formation, aerial hyphal formation, sexual growth, lipid turnover, pigmentation and fungal virulence. Interestingly, ΔFvAtg4 and ΔFvAtg8 were defective in fumonisin B1 (FB1) synthesis, which may have resulted from decreased intracellular levels of alanine in the mutants. Our results indicate that FvAtg4 and FvAtg8 contribute to F. verticillioides pathogenicity by regulating the autophagic pathway to control lipid turnover, fumonisin biosynthesis, and pigmentation during its infectious cycle.

Mycosubtilin Produced by Bacillus subtilis ATCC6633 Inhibits Growth and Mycotoxin Biosynthesis of Fusarium graminearum and Fusarium verticillioides.

Fusarium graminearum and Fusarium verticillioides are fungal pathogens that cause diseases in cereal crops, such as Fusarium head blight (FHB), seedling blight, and stalk rot. They also produce a variety of mycotoxins that reduce crop yields and threaten human and animal health. Several strategies for controlling these diseases have been developed. However, due to a lack of resistant cultivars and the hazards of chemical fungicides, efforts are now focused on the biocontrol of plant diseases, which is a more sustainable and environmentally friendly approach. In the present study, the lipopeptide mycosubtilin purified from Bacillus subtilis ATCC6633 significantly suppressed the growth of F. graminearum PH-1 and F. verticillioides 7600 in vitro. Mycosubtilin caused the destruction and deformation of plasma membranes and cell walls in F. graminearum hyphae. Additionally, mycosubtilin inhibited conidial spore formation and germination of both fungi in a dose-dependent manner. In planta experiments demonstrated the ability of mycosubtilin to control the adverse effects caused by F. graminearum and F. verticillioides on wheat heads and maize kernels, respectively. Mycosubtilin significantly decreased the production of deoxynivalenol (DON) and B-series fumonisins (FB1, FB2 and FB3) in infected grains, with inhibition rates of 48.92, 48.48, 52.42, and 59.44%, respectively. The qRT-PCR analysis showed that mycosubtilin significantly downregulated genes involved in mycotoxin biosynthesis. In conclusion, mycosubtilin produced by B. subtilis ATCC6633 was shown to have potential as a biological agent to control plant diseases and Fusarium toxin contamination caused by F. graminearum and F. verticillioides.

High Capacitive Energy Storage of Nest-Like Porous Graphene Microspheres Electrode with High Mass Loading.

Nest-like porous graphene microspheres (NPGMs) are grown by using a chemical vapor deposition (CVD) method in a fluidized bed reactor from methane and basic magnesium carbonate microspheres (synthesized by a stirring-induced crystallization approach) as carbon source and template, respectively. The CVD-derived NPGMs have a few-layer structure and high electrical conductivity, as well as a three-dimensional individual macroarchitecture accompanied with well-developed pore channels and great structural integrity. As the electrode for a symmetric supercapacitor, the effect of different mass loadings for NPGMs-based electrodes on the capacitive energy-storage performance is investigated. Superior electrochemical properties with respect to gravimetric, areal, and total capacitances, rate capability, and durability are shown by the NPGMs-based symmetric supercapacitors, even at mass loadings up to 10 mg cm-2 . Moreover, the electrochemical behavior of the NPGMs-based electrode is much superior to those of two-dimensional lamella-like graphene and commercial activated carbon.

Fengycin Produced by Bacillus amyloliquefaciens FZB42 Inhibits Fusarium graminearum Growth and Mycotoxins Biosynthesis.

Fusarium graminearum is a notorious pathogen that causes Fusarium head blight (FHB) in cereal crops. It produces secondary metabolites, such as deoxynivalenol, diminishing grain quality and leading to lesser crop yield. Many strategies have been developed to combat this pathogenic fungus; however, considering the lack of resistant cultivars and likelihood of environmental hazards upon using chemical pesticides, efforts have shifted toward the biocontrol of plant diseases, which is a sustainable and eco-friendly approach. Fengycin, derived from Bacillus amyloliquefaciens FZB42, was purified from the crude extract by HPLC and further analyzed by MALDI-TOF-MS. Its application resulted in structural deformations in fungal hyphae, as observed via scanning electron microscopy. In planta experiment revealed the ability of fengycin to suppress F. graminearum growth and highlighted its capacity to combat disease incidence. Fengycin significantly suppressed F. graminearum, and also reduced the deoxynivalenol (DON), 3-acetyldeoxynivalenol (3-ADON), 15-acetyldeoxynivalenol (15-ADON), and zearalenone (ZEN) production in infected grains. To conclude, we report that fengycin produced by B. amyloliquefaciens FZB42 has potential as a biocontrol agent against F. graminearum and can also inhibit the mycotoxins produced by this fungus.