CorNet: Autonomous feature learning in raw Corvis ST data for keratoconus diagnosis via residual CNN approach.

PURPOSE: To ascertain whether the integration of raw Corvis ST data with an end-to-end CNN can enhance the diagnosis of keratoconus (KC). METHOD: The Corvis ST is a non-contact device for in vivo measurement of corneal biomechanics. The CorNet was trained and validated on a dataset consisting of 1786 Corvis ST raw data from 1112 normal eyes and 674 KC eyes. Each raw data consists of the anterior and posterior corneal surface elevation during air-puff induced dynamic deformation. The architecture of CorNet utilizes four ResNet-inspired convolutional structures that employ 1 × 1 convolution in identity mapping. Gradient-weighted Class Activation Mapping (Grad-CAM) was adopted to visualize the attention allocation to diagnostic areas. Discriminative performance was assessed using metrics including the AUC of ROC curve, sensitivity, specificity, precision, accuracy, and F1 score. RESULTS: CorNet demonstrated outstanding performance in distinguishing KC from normal eyes, achieving an AUC of 0.971 (sensitivity: 92.49%, specificity: 91.54%) in the validation set, outperforming the best existing Corvis ST parameters, namely the Corvis Biomechanical Index (CBI) with an AUC of 0.947, and its updated version for Chinese populations (cCBI) with an AUC of 0.963. Though the ROC curve analysis showed no significant difference between CorNet and cCBI (p = 0.295), it indicated a notable difference between CorNet and CBI (p = 0.011). The Grad-CAM visualizations highlighted the significance of corneal deformation data during the loading phase rather than the unloading phase for KC diagnosis. CONCLUSION: This study proposed an end-to-end CNN approach utilizing raw biomechanical data by Corvis ST for KC detection, showing effectiveness comparable to or surpassing existing parameters provided by Corvis ST. The CorNet, autonomously learning comprehensive temporal and spatial features, demonstrated a promising performance for advancing KC diagnosis in ophthalmology.

Comparative analysis of models in predicting the effects of SNPs on TF-DNA binding using large-scale in vitro and in vivo data.

Non-coding variants associated with complex traits can alter the motifs of transcription factor (TF)-deoxyribonucleic acid binding. Although many computational models have been developed to predict the effects of non-coding variants on TF binding, their predictive power lacks systematic evaluation. Here we have evaluated 14 different models built on position weight matrices (PWMs), support vector machines, ordinary least squares and deep neural networks (DNNs), using large-scale in vitro (i.e. SNP-SELEX) and in vivo (i.e. allele-specific binding, ASB) TF binding data. Our results show that the accuracy of each model in predicting SNP effects in vitro significantly exceeds that achieved in vivo. For in vitro variant impact prediction, kmer/gkm-based machine learning methods (deltaSVM_HT-SELEX, QBiC-Pred) trained on in vitro datasets exhibit the best performance. For in vivo ASB variant prediction, DNN-based multitask models (DeepSEA, Sei, Enformer) trained on the ChIP-seq dataset exhibit relatively superior performance. Among the PWM-based methods, tRap demonstrates better performance in both in vitro and in vivo evaluations. In addition, we find that TF classes such as basic leucine zipper factors could be predicted more accurately, whereas those such as C2H2 zinc finger factors are predicted less accurately, aligning with the evolutionary conservation of these TF classes. We also underscore the significance of non-sequence factors such as cis-regulatory element type, TF expression, interactions and post-translational modifications in influencing the in vivo predictive performance of TFs. Our research provides valuable insights into selecting prioritization methods for non-coding variants and further optimizing such models.

Self-cultivating anammox granules for enhancing wastewater nitrogen removal in nitrification-denitrification flocculent sludge system.

The feasibility of self-cultivating anammox granules for enhancing wastewater nitrogen removal was investigated in a nitrification-denitrification flocculent sludge system. Desirable nitrogen removal efficiency of 84 ± 4 % was obtained for the influent carbon to nitrogen ratio of 1-1.3 (NH4+-N: 150-200 mg N/L) via alternate anaerobic/oxic/anoxic mode. Meanwhile, some red granular sludge was formed in the system. The abundance and activity of anaerobic ammonia oxidation bacteria (AnAOB) increased from 'not detected' in seed sludge to 0.57 % and 29.4 ± 0.7 mg N/(g mixed liquor volatile suspended solids·h) in granules, respectively, suggesting successful cultivation of anammox granules. Furthermore, some denitrifying bacteria with capability of partial denitrification were enriched, such as Candidatus Competibacter (2.45 %) and Thauera (5.75 %), which could cooperate with AnAOB, facilitating AnAOB enrichment. Anammox was dominant in nitrogen removal with the contribution to nitrogen removed above 68.8 ± 0.3 %. The strategy of self-cultivating anammox granules could promote the application of anammox.

The Plant Volatile-Sensing Mechanism of Insects and Its Utilization.

Plants and insects are engaged in a tight relationship, with phytophagous insects often utilizing volatile organic substances released by host plants to find food and egg-laying sites. Using plant volatiles as attractants for integrated pest management is vital due to its high efficacy and low environmental toxicity. Using naturally occurring plant volatiles combined with insect olfactory mechanisms to select volatile molecules for screening has proved an effective method for developing plant volatile-based attractant technologies. However, the widespread adoption of this technique is still limited by the lack of a complete understanding of molecular insect olfactory pathways. This paper first describes the nature of plant volatiles and the mechanisms of plant volatile perception by insects. Then, the attraction mechanism of plant volatiles to insects is introduced with the example of Cnaphalocrocis medinalis. Next, the progress of the development and utilization of plant volatiles to manage pests is presented. Finally, the functions played by the olfactory system of insects in recognizing plant volatiles and the application prospects of utilizing volatiles for green pest control are discussed. Understanding the sensing mechanism of insects to plant volatiles and its utilization will be critical for pest management in agriculture.

Increasing carbon to nitrogen ratio promoted anaerobic ammonia-oxidizing bacterial enrichment and advanced nitrogen removal in mainstream anammox system.

The effects of fluctuating organic carbon to nitrogen (C/N) ratios on mainstream simultaneous partial nitrification, anammox, and denitrification (SNAD) process were studied over 376-day period. The nitrogen removal efficiency decreased from 85.0 ± 6.6 % to 75.8 ± 2.8 % as C/N ratio decreased (3.4 â†’ 1.7), but increased to 82.0 ± 1.9 % when C/N ratio raised to 2.9 and to 78.4 ± 3.0 % when C/N ratio decreased again (2.9 â†’ 2.1), indicating that high C/N ratios promoted nitrogen removal. As C/N ratio raised (1.7 â†’ 2.9), anaerobic ammonia-oxidizing bacteria (AnAOB) abundance increased from 1.3 × 109 to 2.0 × 109 copies/L, which explained the improved nitrogen removal. With an elevated C/N ratio, partial nitrification and endogenous partial denitrification reactions were enhanced, providing more nitrite for AnAOB. Additionally, the aerobic_chemoheterotrophy function and particle sizes increased, forming more stable anoxic microenvironment for AnAOB. Overall, increasing C/N ratio promoted the stability of mainstream SNAD.

Metabolome profiling and transcriptome analysis filling the early crucial missing steps of piperine biosynthesis in Piper nigrum L.

Black pepper (Piper nigrum L.), the world renown as the King of Spices, is not only a flavorsome spice but also a traditional herb. Piperine, a species-specific piper amide, is responsible for the major bioactivity and pungent flavor of black pepper. However, several key steps for the biosynthesis of piperoyl-CoA (acyl-donor) and piperidine (acyl-acceptor), two direct precursors for piperine, remain unknown. In this study, we used guilt-by-association analysis of the combined metabolome and transcriptome, to identify two feruloyldiketide-CoA synthases responsible for the production of the C5 side chain scaffold feruloyldiketide-CoA intermediate, which is considered the first and important step to branch metabolic fluxes from phenylpropanoid pathway to piperine biosynthesis. In addition, we also identified the first two key enzymes for piperidine biosynthesis derived from lysine in P. nigrum, namely a lysine decarboxylase and a copper amine oxidase. These enzymes catalyze the production of cadaverine and 1-piperideine, the precursors of piperidine. In vivo and in vitro experiments verified the catalytic capability of them. In conclusion, our findings revealed enigmatic key steps of piperine biosynthetic pathway and thus provide a powerful reference for dissecting the biosynthetic logic of other piper amides.

Migration of microorganisms between nitrification-denitrification flocs, anammox biofilms and blank carriers during mainstream anammox start-up.

To solve the shortage of inoculum, the feasibility of establishing simultaneous partial nitrification, anammox, and denitrification (SNAD) reactor through inoculating nitrification-denitrification sludge, anammox biofilm and blank carriers was investigated. Advanced nitrogen removal efficiency of 91.2 ± 3.6 % was achieved. Bacteria related to nitrogen removal and fermentation were enriched in anammox biofilm, blank carriers and flocs, and the abundance of dominant anaerobic ammonia oxidizing bacteria (AnAOB), Candidatus Brocadia, reached 3.4 %, 0.5 % and 0.3 %, respectively. Candidatus Competibacter and Calorithrix became the dominant denitrifying bacteria (DNB) and fermentative bacteria (FB), respectively. The SNAD system was successfully established, and new mature biofilms formed in blank carriers, which could provide inoculum for other anammox processes. Partial nitrification, partial denitrification and aerobic_chemoheterotrophy were existed and facilitated AnAOB enrichment. Microbial correlation networks revealed the cooperation between DNB, FB and AnAOB that promoted nitrogen removal. Overall, the SNAD process was started up through inoculating more accessible inoculum.

Performance of Corvis ST Parameters Including Updated Stress-Strain Index in Differentiating Between Normal, Forme-Fruste, Subclinical, and Clinical Keratoconic Eyes.

PURPOSE: This study seeks to evaluate the ability of the updated stress strain index (SSIv2) and other Corvis ST biomechanical parameters in distinguishing between keratoconus at different disease stages and normal eyes. DESIGN: Diagnostic accuracy analysis to distinguish disease stages. METHODS: 1084 eyes were included and divided into groups of normal (199 eyes), forme fruste keratoconus (FFKC, 194 eyes), subclinical keratoconus (SKC, 113 eyes), mild clinical keratoconus (CKC-Ⅰ, 175 eyes), moderate clinical keratoconus (CKC-Ⅱ, 204 eyes), and severe clinical keratoconus (CKC-Ⅲ, 199 eyes). Each eye was subjected to a Corvis ST examination to determine the central corneal thickness (CCT), biomechanically corrected intraocular pressure (bIOP), SSIv2 (updated stress-strain index), and other 8 Corvis parameters including the stress-strain index (SSIv1), stiffness parameter at first applanation (SP-A1), first applanation time (A1T), Ambrósio relational thickness to the horizontal profile (ARTh), integrated inverse radius (IIR), maximum deformation amplitude (DAM), ratio between deformation amplitude at the apex and at 2 mm nasal and temporal (DARatio2), and Corvis biomechanical index (CBI). The sensitivity and specificity of these parameters in diagnosing keratoconus were analyzed through receiver operating characteristic curves. RESULTS: Before and after correction for CCT and bIOP, SSIv2 and ARTh were significantly higher and IIR and CBI were significantly lower in the normal group than in the FFKC group, SKC group and the 3 CKC groups (all P < .05). There were also significant correlations between the values of SSIv2, ARTh, IIR, CBI, and the CKC severity (all P < .05). AUC of SSIv2 was significantly higher than all other Corvis parameters in distinguishing normal eyes from FFKC, followed by IIR, ARTh and CBI. CONCLUSION: Corvis ST's updated stress-strain index, SSIv2, demonstrated superior performance in differentiating between normal and keratoconic corneas, and between corneas with different keratoconus stages. Similar, but less pronounced, performance was demonstrated by the IIR, ARTh and CBI.

Achieving advanced nitrogen removal and excess sludge treatment via single nitritation/anammox-fermentation combined system.

The feasibility of treating wastewater and excess sludge via simultaneous nitritation, anammox, denitrification and fermentation (SNADF) was investigated in three parallel sequencing batch reactors (SBRs). SBR2 and SBR3 received exogenous nitrification-denitrification sludge and thermal hydrolysis sludge, respectively. Nitrogen removal efficiencies of 92.8 ± 5.9%, 94.6 ± 4.1%, 93.4 ± 4.8% were achieved in SBR1, SBR2, and SBR3, respectively (influent ammonium: 56.0-74.0 mg N/L), with low observed sludge yield of 0.02-0.15, -0.06-0.11, -0.17-0.05 kg mixed liquor suspended solids (MLSS)/kg chemical oxygen demand (COD). Anammox bacterial abundances increased from 3.6 × 109 ± 2.8 × 108 to 8.1 × 109 ± 2.3 × 108, 1.5 × 1010 ± 1.1 × 108, and 1.4 × 1010 ± 2.9 × 108 copies/L in SBR1-SBR3, respectively. The abundances of Nitrosomonas, genes (amo, hao) for partial nitrification, and narGHI genes (nitrate â†’ nitrite) in dominant partial denitrifying bacteria (Candidatus Competibacter) were higher in SBR2 and SBR3 than that in SBR1. These results suggested that adding excess sludge promoted sludge reduction, nitrite production and anammox bacterial enrichment. The SNADF system could treat excess sludge, meanwhile, achieve advanced nitrogen removal.

Increasing specific biomass nitrogen load facilitated nitrite accumulation in mainstream wastewater treatment.

By regulating influent nitrogen loading rate (NLR) and solids retention time (SRT), the effect of specific biomass nitrogen load (BNL) on the start-up of mainstream partial nitrification (PN) was investigated in five parallel sequencing batch reactors inoculated with ordinary nitrification sludge. The results showed that increasing BNL by both methods could achieve nitrite accumulation. Moreover, a high initial activity of ammonium oxidizing bacteria (AOB) accelerated nitrite accumulation. Increasing influent NLR (ammonium: 55-70 mg N/L) achieved only 30%-40% of nitrite accumulation ratio (NAR) and gradually decreased with reactor operation. By increasing BNL via controlling SRT (30 days), desirable PN with an average NAR of 81.7 ± 4.4% (effluent nitrite: ∼10 mg N/L) was obtained. Nitrite oxidizing bacteria (NOB) were effectively inhibited, and the AOB to NOB activity ratio increased from 1.5 to 7.8, promoting efficient nitrite accumulation. Overall, increasing BNL by regulating SRT was a potential method for start-up of mainstream PN.

Enhancing nitrite production rate made anammox bacteria have a competitive advantage over nitrite oxidizing bacteria in mainstream anammox system.

Nitrite oxidizing bacteria (NOB) is easy to accumulate in the mainstream anammox process, leading to the decrease of anammox bacterial abundance and deterioration of nitrogen removal. In this study, anammox bacteria was gradually enriched by increasing nitrite production rate under intermittent aeration despite high NOB abundance. With the DO increased from 0.4 to 0.6 mg/L, Nitrosomonas increased from 0.14% to 0.22%, providing more nitrite for anammox bacteria and promoting its enrichment (grew by 77.4%). Adding extra nitrite of 7.14 mg N/(L·h) during the aeration phase to reactor could further increase anammox bacterial abundance by 117.6%, which was higher than the control reactor (40.2%). In contrast, NOB abundance decreased from 1.4 × 1010 to 1.2 × 1010 copies/L. The results suggested that anammox bacteria had a competitive advantage for nitrite over NOB with increasing nitrite production rate. In addition, Thauera and Dechloromonas, which were responsible for reducing nitrate to nitrite, provided additional substrates for anammox bacteria. Overall, this research provides a new idea for mainstream anammox applications. PRACTITIONER POINTS: Inhibiting NOB might be no longer necessary and difficult for mainstream anammox. Anammox bacteria competed for more nitrite with NOB when nitrite production rate increased. Increasing DO from 0.4 to 0.6 mg/L facilitated anammox bacterial growth and nitrogen removal.

Paravertebral block vs. epidural block for percutaneous nephrolithotomy: A prospective, randomized study.

Background: Percutaneous nephrolithotripsy (PCNL) is the main method for urinary calculi. An anesthesia method with little effect on the blood circulation and which does not affect the postoperative activity of a patient is lacking. Objective: To compare the effects of paravertebral nerve block (PNB) and epidural block (EPB) on quadriceps femoris muscle (QFM) strength in patients after PCNL. Methods: 163 patients were separated into two groups: EPB (81) and PNB (82). Primary outcome parameters were QFM strength and range of motion (RoM) of the knee 1 h, 2 h, 3 h, and 24 h after anesthesia induction (AI). Secondary outcome parameters were: time from AI beginning to first ambulation; time of sensory-plane recovery; amount of additional analgesics given during and after surgery; prevalence of nausea and vomiting; duration of hospital stay (DoHS); mean arterial pressure (MAP), heart rate (HR), and oxygen saturation (SpO2) before, 0.5 h, and 1 h after AI; visual analog scale (VAS) score 0.5 h, 1 h, 2 h, 3 h and 24 h after AI. Results: There was no significant difference in QFM strength or knee RoM before or 24 h after AI between the two groups (P > 0.05). The time from AI to first ambulation was shorter (P < 0.05) and the sensory plane took longer to recover (P < 0.05) in the PNB group than in the EPB group. The amount of additional analgesics during surgery was more in the PNB group than in the EPB group (P < 0.05), but there was no significant difference after surgery (P > 0.05). VAS scores were higher in the PNB group than in the EPB group 0.5 after AI (P < 0.05). MAP 1 h after AI was higher in the PNB group than in the EPB group (P < 0.05). There was no significant difference in the prevalence of postoperative nausea and vomiting, DoHS, HR, or SpO2 at 0.5 h and 1 h after AI between the two groups (P > 0.05). Conclusions: For patients undergoing PCNL, PNB can meet the need for surgical analgesia while having little effect on QFM strength. Trial registration: http://www.chictr.org.cn/, identifier ChiCTR2200060606.

Spin Hall effect from bipolaron dynamics in organics.

Using an extended Su-Schrieffer-Heeger model and a nonadiabatic dynamics method, we investigate the dynamics of bipolarons in coupled nondegenerate organic chains including the spin-orbit coupling and interchain coupling. By tracing the time-dependent evolution of the charges and spins in each chain, an obvious oscillating spin Hall effect (SHE) from the bipolaron transport is revealed. The results are compared with that from polaron-dominated transport. A reduction of amplitude and an increase of oscillating frequency are observed for the SHE from the bipolaron transport. The mechanism is attributed to the enhanced skew scattering off the larger transient deformations of the chains in the case of the bipolaron. Spectrum analysis by fast Fourier transform of the SHE signal demonstrates a distinct shift of two characteristic peaks to a higher onset frequency compared to the polaron transport. The charge-spin conversion efficiency is also compared, where a larger conversion efficiency is obtained from the bipolaron transport due to the lower saturated velocity. The effects of the strength of the electric field and the interactions are discussed. This work reveals the role of the bipolaron in organic SHE and provides a feasible way to achieve larger conversion efficiency by controlling the species of carriers with the concentration of the dopant.

Dynamic topography analysis of the cornea and its application to the diagnosis of keratoconus.

PROPOSE: To establish a dynamic topography analysis method which simulates the dynamic biomechanical response of the cornea and reveals the variations of such response within the corneal surface, and thereafter to propose and clinically evaluate new parameters for the definite diagnosis of keratoconus. METHODS: 58 normal (Normal) and 56 keratoconus (KC) subjects were retrospectively included. Personalized corneal air-puff model was established using corneal topography data by Pentacam for each subject, and the dynamic deformation under air-puff loading was simulated using finite element method, which then enabled calculations of corneal biomechanical parameters of the entire corneal surface along any meridian. Variations in these parameters across different meridians and between different groups were explored by two-way repeated measurement analysis of variance. New dynamic topography parameters were proposed as the range of the calculated biomechanical parameters within the entire corneal surface, and the AUC of ROC curve was used to compare the diagnostic efficiency of newly proposed and existing parameters. RESULTS: Corneal biomechanical parameters measured in different meridians varied significantly which were more pronounced in KC group due to its irregularity in corneal morphology. Considering such between-meridian variations thus led to improved diagnostic efficiency of KC as presented by the proposed dynamic topography parameter rIR with an AUC of 0.992 (sensitivity: 91.1%, specificity: 100%), significantly better than the current topography and biomechanical parameters. CONCLUSIONS: The diagnosis of keratoconus may be affected by the significant variations of corneal biomechanical parameters due to corneal morphology irregularity. By considering such variations, the current study established the dynamic topography analysis process which benefits from the high accuracy of (static) corneal topography measurement while improving its diagnosis capacity. The proposed dynamic topography parameters, especially the rIR parameter, showed comparable or better diagnostic efficiency for KC than existing topography and biomechanical parameters, which can be of great clinical significance for clinics without access to instrument for biomechanical evaluations.

Cellulose nanocrystals-based bio-composite optical materials for reversible colorimetric responsive films and coatings.

Photonic materials with a tunable chiral nematic structure that can selectively reflect light dynamically are valuable for applications in smart responsive materials. Here, we prepared potential photonic composites with a chiral nematic structure by forming cellulose nanocrystals (CNCs) and waterborne polyurethane (WPU) composites with different compositions on different substrates by evaporation-induced self-assembly. With increasing WPU content, the reflected wavelength increased from 400 to 680 nm, which was mainly caused by the increase of the chiral nematic pitch. In addition, the mechanical properties were better for higher WPU content. WPU was sensitive to small amounts of moisture in ethanol owing to the swollen WPU after absorbing water will increase the helical pitch. The reversible red shift induced by moisture was approximately 100 nm. When wood was used as the substrate, the CNCs still self-assembled to form chiral nematic structures and the adhesion forces of the composites to the wood substrate were strong. By using MgCl2 solution as an ink, invisible patterns can be written on the coating, which can be revealed temporarily by ethanol. In addition, the invisible pattern of photonic coating is rewritable. The easily prepared environmentally friendly photonic composite has great potential in sensors, anti-counterfeiting labels and smart decorative coatings.

A novel simultaneous partial nitrification, anammox, denitrification and fermentation process: Enhancing nitrogen removal and sludge reduction in a single reactor.

This study verified the feasibility of simultaneous partial nitrification, anammox, denitrification and fermentation process under intermittent aeration in a single reactor, and explored the impact of dissolved oxygen (DO) on the synergy between fermentation and nitrogen removal. An advanced nitrogen removal efficiency of 92.8 % and a low observed sludge yield of 0.0268-0.1474 kgMLSS/kgCOD were achieved. In-situ test showed that nitrate and ammonium decreased synchronously in the absence of organic matter, indicating the possibility of simultaneous partial denitrification, anammox and fermentation. Additionally, the abundance of functional genes for acetate production was 66,894 hits, while the key genes relevant to methanogenesis were only 348 hits, which suggested that fermentation might stop at the acid-producing stage and promote partial denitrification-anammox reaction, achieving simultaneous sludge reduction and advanced nitrogen removal performance. When DO increased from 0.1-0.3 to 0.4-0.6 mg/L, the nitrogen removal efficiency was increased (63.9 %→92.8 %) while sludge reduction was negatively affected.

Disease resistance conferred by components of essential chrysanthemum oil and the epigenetic regulation of OsTPS1.

The sesquiterpene alpha-bisabolol is the predominant active ingredient in essential oils that are highly valued in the cosmetics industry due to its wound healing, anti-inflammatory, and skin-soothing properties. Alpha-bisabolol was thought to be restricted to Compositae plants. Here we reveal that alpha-bisabolol is also synthesized in rice, a non-Compositae plant, where it acts as a novel sesquiterpene phytoalexin. Overexpressing the gene responsible for the biosynthesis of alpha-bisabolol, OsTPS1, conferred bacterial blight resistance in rice. Phylogenomic analyses revealed that alpha-bisabolol-synthesizing enzymes in rice and Compositae evolved independently. Further experiments demonstrated that the natural variation in the disease resistance level was associated with differential transcription of OsTPS1 due to polymorphisms in its promoter. We demonstrated that OsTPS1 was regulated at the epigenetic level by JMJ705 through the methyl jasmonate pathway. These data reveal the cross-family accumulation and regulatory mechanisms of alpha-bisabolol production.

Mechanically Enhanced Nanocrystalline Cellulose/Reduced Graphene Oxide/Polyethylene Glycol Electrically Conductive Composite Film.

Traditional conductive materials do not meet the increasing requirements of electronic products because of such materials' high rigidity, poor flexibility, and slow biodegradation after disposal. Preparing flexible conductive materials with excellent mechanical properties is an active area of research. The key to flexible conductive materials lies in the combination of the polymer matrix and conductive components. This combination can be achieved by making a film of renewable nano-microcrystalline cellulose (NCC) and reduced graphene oxide (rGO) with excellent electrical conductivity-by simple filtration and introducing polyethylene glycol (PEG) to enhance the functionality of the composite film. Graphene imparted conductivity to the composite film, which reached 5.67 S·m-1. A reinforced NCC/rGO/PEG-4 composite film with a thickness of only 21 µm exhibited a tensile strength of 30.56 MPa, which was 83% higher than that of the sample without PEG (16.71 MPa), and toughness of 727.18 kJ·m-3, which was about 132% higher than that of the control sample (NCC/rGO, 313.86 kJ·m-3). This ultra-thin conductive composite film-which can be prepared simply, consists of environmentally sustainable and biodegradable raw materials, and exhibits excellent mechanical properties-has substantial potential for applications in e.g., flexible electronic wearable devices, electrodes, and capacitors.

Genome-wide association studies of five free amino acid levels in rice.

Rice (Oryza sativa L.) is one of the important staple foods for human consumption and livestock use. As a complex quality trait, free amino acid (FAA) content in rice is of nutritional importance. To dissect the genetic mechanism of FAA level, five amino acids' (Val, Leu, Ile, Arg, and Trp) content and 4,325,832 high-quality SNPs of 448 rice accessions were used to conduct genome-wide association studies (GWAS) with nine different methods. Of these methods, one single-locus method (GEMMA), seven multi-locus methods (mrMLM, pLARmEB, FASTmrEMMA, pKWmEB, FASTmrMLM, ISIS EM-BLASSO, and FarmCPU), and the recent released 3VmrMLM were adopted for methodological comparison of quantitative trait nucleotide (QTN) detection and identification of stable quantitative trait nucleotide loci (QTLs). As a result, 987 QTNs were identified by eight multi-locus GWAS methods; FASTmrEMMA detected the most QTNs (245), followed by 3VmrMLM (160), and GEMMA detected the least QTNs (0). Among 88 stable QTLs identified by the above methods, 3VmrMLM has some advantages, such as the most common QTNs, the highest LOD score, and the highest proportion of all detected stable QTLs. Around these stable QTLs, candidate genes were found in the GO classification to be involved in the primary metabolic process, biosynthetic process, and catalytic activity, and shown in KEGG analysis to have participated in metabolic pathways, biosynthesis of amino acids, and tryptophan metabolism. Natural variations of candidate genes resulting in the content alteration of five FAAs were identified in this association panel. In addition, 95 QTN-by-environment interactions (QEIs) of five FAA levels were detected by 3VmrMLM only. GO classification showed that the candidate genes got involved in the primary metabolic process, transport, and catalytic activity. Candidate genes of QEIs played important roles in valine, leucine, and isoleucine degradation (QEI_09_03978551 and candidate gene LOC_Os09g07830 in the Leu dataset), tryptophan metabolism (QEI_01_00617184 and candidate gene LOC_Os01g02020 in the Trp dataset), and glutathione metabolism (QEI_12_09153839 and candidate gene LOC_Os12g16200 in the Arg dataset) pathways through KEGG analysis. As an alternative of the multi-locus GWAS method, these findings suggested that the application of 3VmrMLM may provide new insights into better understanding FAA accumulation and facilitate the molecular breeding of rice with high FAA level.