A highly sensitive point-of-care detection platform for Salmonella typhimurium by integrating magnetic enrichment and fluorescent CsPbBr3@SiO2.

A platform was designed based on Fe3O4 and CsPbBr3@SiO2 for integrated magnetic enrichment-fluorescence detection of Salmonella typhimurium, which significantly simplifies the detection process and enhances the working efficiency. Fe3O4 served as a magnetic enrichment unit for the capture of S. typhimurium. CsPbBr3@SiO2 was employed as a fluorescence-sensing unit for quantitative signal output, where SiO2 was introduced to strengthen the stability of CsPbBr3, improve its biomodificability, and prevent lead leakage. More importantly, the SiO2 shell shows neglectable absorption or scattering towards fluorescence, making the CsPbBr3@SiO2 exhibit a high quantum yield of 74.4%. After magnetic enrichment, the decreasing rate of the fluorescence emission intensity of the CsPbBr3@SiO2 supernatant at 527 nm under excitation light at UV 365 nm showed a strong linear correlation with S. typhimurium concentration of 1 × 102~1 × 108 CFU∙mL-1, and the limit of detection (LOD) reached 12.72 CFU∙mL-1. This platform has demonstrated outstanding stability, reproducibility, and resistance to interference, which provides an alternative for convenient and quantitative detection of S. typhimurium.

Active vision in freely moving marmosets using head-mounted eye tracking.

Our understanding of how vision functions as primates actively navigate the real-world is remarkably sparse. As most data have been limited to chaired and typically head-restrained animals, the synergistic interactions of different motor actions/plans inherent to active sensing - e.g. eyes, head, posture, movement, etc. - on visual perception are largely unknown. To address this considerable gap in knowledge, we developed an innovative wireless head-mounted eye tracking system called CEREBRO for small mammals, such as marmoset monkeys. Our system performs C hair-free E ye- Re cording using B ackpack mounted mic RO controllers. Because eye illumination and environment lighting change continuously in natural contexts, we developed a segmentation artificial neural network to perform robust pupil tracking in these conditions. Leveraging this innovative system to investigate active vision, we demonstrate that although freely-moving marmosets exhibit frequent compensatory eye movements equivalent to other primates, including humans, the predictability of the visual system is enhanced when animals are freely-moving relative to when they are head-fixed. Moreover, despite increases in eye/head-motion during locomotion, gaze stabilization actually improved over periods when the monkeys were stationary. Rather than impair vision, the dynamics of gaze stabilization in freely-moving primates has been optimized over evolution to enable active sensing during natural exploration.

Modeling the carbon dynamics of ecosystem in a typical permafrost area.

Climate change poses mounting threats to fragile alpine ecosystem worldwide. Quantifying changes in carbon stocks in response to the shifting climate was important for developing climate change mitigation and adaptation strategies. This study utilized a process-based land model (Community Land Model 5.0) to analyze spatiotemporal variations in vegetation carbon stock (VCS) and soil organic carbon stock (SOCS) across a typical permafrost area - Qinghai Province, China, from 2000 to 2018. Multiple potential factors influencing carbon stocks dynamics were analyzed, including climate, vegetation, soil hydrothermal status, and soil properties. The results indicated that provincial vegetation carbon storage was 0.22 PgC (0.32 kg/m2) and soil organic carbon pool was 9.12 PgC (13.03 kg/m2). VCS showed a mild increase while SOCS exhibited fluctuating uptrends during this period. Higher carbon stocks were observed in forest (21.74 kg/m2) and alpine meadow (18.08 kg/m2) compared to alpine steppes (9.63 kg/m2). Over 90 % of the carbon was stored in the 0-30 cm topsoil layer. The contribution rates of soil carbon in the 30-60 cm and 60-100 cm soil layers were significantly small, despite increasing stocks across all depths. Solar radiation, temperature, and NDVI emerged as primary influential factors for overall carbon stocks, exhibiting noticeable spatial variability. For SOCS at different depths, the normalized differential vegetation index (NDVI) was the foremost predictor of landscape-level carbon distributions, which explained 52.8 % of SOCS variability in shallow layers (0-30 cm) but dropped to just 12.97 % at the depth of 30-60 cm. However, the dominance of NDVI diminished along the soil depth gradients, superseded by radiation and precipitation. Additionally, with an increase in soil depth, the influence of inherent soil properties also increased. This simulation provided crucial insights for landscape-scale carbon responses to climate change, and offered valuable reference for other climate change-sensitive areas in terms of ecosystem carbon management.

First-principles calculation on electronic properties of hydrogen evolution reaction of Ni-based electrode surfaces with different monatomic doping.

At present, the hydrogen evolution reaction (HER) of Ni-based electrode has an important influence on water electrolysis hydrogen production technology, involving complex electrochemical process of electrode. In this project, Materials Studio (MS) software was used to design and construct Ni-based electrode surface (NES) models with monatomic Mo, Co, Fe, Cr doping, and the NES models attached 1 H atom and 2H atoms were denoted as the NES-H models and NES-2H model, respectively. Then the first-principles calculation was carried out. The results showed that the doping of different atoms can effectively change the work function of the pure Ni. In the charge transfer process of the four NES-2H models, the distance between the two H atoms is most affected by Mo doping, and they leave the Ni electrode surface as a single H ion, respectively, while the effect on Co, Fe and Cr doping is relatively consistent, and they leave the Ni electrode surface with H2 molecules, respectively. The doping of four single atoms changes the distance of valence band (VB) top and conduction band (CB) bottom from Fermi level in NES, NES-H and NES-2H models, and affects the HER, in which Mo doping has the greatest effect. The TDOS of the above models is mainly derived from the PDOS of the d orbitals of the doped atoms and Ni atoms. The results will provide a theoretical basis for the research and development of Ni-based electrode materials in HER.

Representing the dynamics of natural marmoset vocal behaviors in frontal cortex.

Here we tested the respective contributions of primate premotor and prefrontal cortex to support vocal behavior. We applied a model-based GLM analysis that better accounts for the inherent variance in natural, continuous behaviors to characterize the activity of neurons throughout frontal cortex as freely-moving marmosets engaged in conversational exchanges. While analyses revealed functional clusters of neural activity related to the different processes involved in the vocal behavior, these clusters did not map to subfields of prefrontal or premotor cortex, as has been observed in more conventional task-based paradigms. Our results suggest a distributed functional organization for the myriad neural mechanisms underlying natural social interactions and has implications for our concepts of the role that frontal cortex plays in governing ethological behaviors in primates.

Interaction Between SARS-CoV-2 Spike Protein S1 Subunit and Oyster Heat Shock Protein 70.

There is growing evidence that severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) contaminates the marine environment and is bioaccumulated in filter-feeding shellfish. Previous study shows the Pacific oyster tissues can bioaccumulate the SARS-CoV-2, and the oyster heat shock protein 70 (oHSP70) may play as the primary attachment receptor to bind SARS-CoV-2's recombinant spike protein S1 subunit (rS1). However, detailed information about the interaction between rS1 and oHSP70 is still unknown. In this study, we confirmed that the affinity of recombinant oHSP70 (roHSP70) for rS1 (KD = 20.4 nM) is comparable to the receptor-binding affinity of rACE2 for rS1 (KD = 16.7 nM) by surface plasmon resonance (SPR)-based Biacore and further validated by enzyme-linked immunosorbent assay (ELISA). Three truncated proteins (roHSP70-N/C/M) and five mutated proteins (p.I229del, p.D457del, p.V491_K495del, p.K556I, and p.ΣroHSP70) were constructed according to the molecular docking results. All three truncated proteins have significantly lower affinity for rS1 than the full-length roHSP70, indicating that all three segments of roHSP70 are involved in binding to rS1. Further, the results of SPR and ELISA showed that all five mutant proteins had significantly lower affinity for rS1 than roHSP70, suggesting that amino acids at these sites are involved in binding to rS1. This study provides a preliminary theoretical basis for the bioaccumulation of SARS-CoV-2 in oyster tissues or using roHSP70 as the capture unit to selectively enrich virus particles for detection.

[A novel periodontal endoscopy-aided non-incisional periodontal regeneration technique：a case series study].

PURPOSE: To investigate the effect of endoscopy-aided non-incisional periodontal regeneration technique (NIT) in the treatment of alveolar bone angular resorption. METHODS: Thirteen patients with severe periodontitis(13 diseased teeth) were selected. All patients had alveolar bone angular resorption on adjacent surface. The patients received NIT treatment 6 weeks after periodontal primary therapy. The visualization of subgingival environment was acquired by the periodontal endoscopy. Following the removal of the subgingival plaque, calculus and intra-bony granulation tissue, bone grafting materials were placed into the intra-bony defects with the assistance of a delicate gingival protector. No flap was elevated and no sutures were applied. Probing depth (PD), gingival recession (GR), clinical attachment level (CAL), as well as radiographic parameters were evaluated at baseline and 2 years after treatment. SPSS 22.0 software package was used for data analysis. RESULTS: At 2-years follow-up, an average CAL gain of (3.65±2.10) mm (P＜0.001), PD reduction of (4.42±1.66) mm (P＜0.001), and minimal increase in GR of (0.38±0.87) mm (P=0.25) were observed. Alveolar bone was significantly improved at 2-years follow-up on radiographs (P＜0.001). CONCLUSIONS: For angular resorption site of alveolar bone, NIT treatment can obtain good periodontal regeneration results without flap inversion.

Diagnostic accuracy of artificial intelligence assisted clinical imaging in the detection of oral potentially malignant disorders and oral cancer: A systematic review and meta-analysis.

BACKGROUND: The objective of this study is to examine the application of AI algorithms in detecting OPMD and oral cancerous lesions, and to evaluate the accuracy variations among different imaging tools employed in these diagnostic processes. MATERIALS AND METHODS: A systematic search was conducted in four databases: Embase, Web of Science, PubMed, and Scopus. The inclusion criteria included studies using machine learning algorithms to provide diagnostic information on specific oral lesions, prospective or retrospective design, and inclusion of OPMD. Sensitivity and specificity analyses were also required. Forest plots were generated to display overall diagnostic odds ratio (DOR), sensitivity, specificity, negative predictive values, and summary receiver operating characteristic (SROC) curves. Meta-regression analysis was conducted to examine potential differences among different imaging tools. RESULTS: The overall DOR for AI-based screening of OPMD and oral mucosal cancerous lesions from normal mucosa was 68.438 (95%CI= [39.484, 118.623], I2 = 86%). The area under the SROC curve was 0.938, indicating excellent diagnostic performance. AI-assisted screening showed a sensitivity of 89.9% (95%CI= [0.866,0.925]; I2 = 81%), specificity of 89.2% (95%CI= [0.851,0.922], I2 = 79%), and a high negative predictive value of 89.5% (95%CI= [0.851; 0.927], I2 = 96%). Meta-regression analysis revealed no significant difference among the three image tools. After generating a GOSH plot, the DOR was calculated to be 49.30, and the area under the SROC curve was 0.877. Additionally, sensitivity, specificity, and negative predictive value were 90.5% (95%CI [0.873,0.929], I2=4%), 87.0% (95%CI [0.813,0.912], I2=49%) and 90.1% (95%CI [0.860,0.931], I2=57%), respectively. Subgroup analysis showed that clinical photography had the highest diagnostic accuracy. CONCLUSIONS: AI-based detection using clinical photography shows a high diagnostic odds ratio and is easily accessible in the current era with billions of phone subscribers globally. This indicates that there is significant potential for AI to enhance the diagnostic capabilities of general practitioners to the level of specialists by utilizing clinical photographs, without the need for expensive specialized imaging equipment.

Near-infrared responsive gold nanorods for highly sensitive colorimetric and photothermal lateral flow immuno-detection of SARS-CoV-2.

The highly infectious characteristics of coronavirus disease 2019 (COVID-19), which is caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), highlight the necessity of sensitive and rapid nucleocapsid (N) protein-based antigen testing for early triage and epidemic management. In this study, a colorimetric and photothermal dual-mode lateral flow immunoassay (LFIA) platform for the rapid and sensitive detection of the SARS-CoV-2 N protein was developed based on gold nanorods (GNRs), which possessed tunable local surface plasma resonance (LSPR) absorption peaks from UV-visible to near-infrared (NIR). The LSPR peak was adjusted to match the NIR emission laser 808 nm by controlling the length-to-diameter ratio, which could maximize the photothermal conversion efficiency and achieve photothermal detection signal amplification. Qualitative detection of SARS-CoV-2 N protein was achieved by observing the strip color, and the limit of detection was 2 ng mL-1, while that for photothermal detection was 0.096 ng mL-1. Artificial saliva samples spiked with the N protein were analyzed with the recoveries ranging from 84.38% to 107.72%. The intra-assay and inter-assay coefficients of variation were 6.76% and 10.39%, respectively. We further evaluated the reliability of this platform by detecting 40 clinical samples collected from nasal swabs, and the results matched well with that of nucleic acid detection (87.5%). This method shows great promise in early disease diagnosis and screening.

Circadian rhythm response and its effect on photosynthetic characteristics of the Lhcb family genes in tea plant.

BACKGROUND: The circadian clock, also known as the circadian rhythm, is responsible for predicting daily and seasonal changes in the environment, and adjusting various physiological and developmental processes to the appropriate times during plant growth and development. The circadian clock controls the expression of the Lhcb gene, which encodes the chlorophyll a/b binding protein. However, the roles of the Lhcb gene in tea plant remain unclear. RESULTS: In this study, a total of 16 CsLhcb genes were identified based on the tea plant genome, which were distributed on 8 chromosomes of the tea plant. The promoter regions of CsLhcb genes have a variety of cis-acting elements including hormonal, abiotic stress responses and light response elements. The CsLhcb family genes are involved in the light response process in tea plant. The photosynthetic parameter of tea leaves showed rhythmic changes during the two photoperiod periods (48 h). Stomata are basically open during the day and closed at night. Real-time quantitative PCR results showed that most of the CsLhcb family genes were highly expressed during the day, but were less expressed at night. CONCLUSIONS: Results indicated that CsLhcb genes were involved in the circadian clock process of tea plant, it also provided potential references for further understanding of the function of CsLhcb gene family in tea plant.

Propane-2-sulfonic acid octadec-9-enyl-amide, a novel PPARα/γ dual agonist, attenuates molecular pathological alterations in learning memory in AD mice.

OBJECTIVE: Previous studies have revealed that Propane-2-sulfonic acid octadec-9-enyl-amide(N15) exerts a protective role in the inflammatory response after ischemic stroke and in neuronal damage. However, little is known about N15 in Alzheimer's disease (AD). The aim of this study was to investigate the effects of N15 on AD and explore the underlying molecular mechanism. METHODS: AD mice model was established by lateral ventricular injection with Aß25-35. N15 was daily intraperitoneal administered for 28 days. Morris Water Maze was used to evaluate the neurocognitive function of the mice. The expression of PPARα/γ, brain-derived neurotrophic factor (BDNF), Neurotrophin-3 (NT3), ADAM10, PS1 and BACE1 were measured by qPCR. Aß amyloid in the hippocampus was measured by Congo red assay. Toluidine blue staining was used to detect the neuronal apoptosis. Protein levels of ADAM10, PS1 and BACE1 were determined using immunoblotting. RESULTS: N15 treatment significantly reduced neurocognitive dysfunction, which also significantly activated the expression of PPARα/γ at an optimal dose of 200 mg/kg. Administration of N15 alleviated the formation of Aß amyloid in the hippocampus of AD mice, enhanced the BDNF mRNA expression, decreased the mRNA and protein levels of PS1 and BACE1, upregulated ADAM10 mRNA and protein levels. CONCLUSION: N15 exerts its neuroprotective effects through the activation of PPARα/γ and may be a potential drug for the treatment of AD.

Application of tandem mass spectrometry in the screening and diagnosis of mucopolysaccharidoses.

Mucopolysaccharidoses (MPSs) are caused by a deficiency in the enzymes needed to degrade glycosaminoglycans (GAGs) in the lysosome. The storage of GAGs leads to the involvement of several systems and even to the death of the patient. In recent years, an increasing number of therapies have increased the treatment options available to patients. Early treatment is beneficial in improving the prognosis, but children with MPSs are often delayed in their diagnosis. Therefore, there is an urgent need to develop a method for early screening and diagnosis of the disease. Tandem mass spectrometry (MS/MS) is an analytical method that can detect multiple substrates or enzymes simultaneously. GAGs are reliable markers of MPSs. MS/MS can be used to screen children at an early stage of the disease, to improve prognosis by treating them before symptoms appear, to evaluate the effectiveness of treatment, and for metabolomic analysis or to find suitable biomarkers. In the future, MS/MS could be used to further identify suitable biomarkers for MPSs for early diagnosis and to detect efficacy.

Processing Method and Performance Evaluation of Flame-Retardant Corrugated Sandwich Panel.

In this study, in order to expand the engineering application range of thermoplastic corrugated sheets, flame-retardant thermoplastic corrugated sheets were prepared by the thermoplastic molding method. Based on our previous research results, we prepared flame-retardant prepreg tapes with the flame retardant addition accounting for 15%, 20%, and 25% of the resin matrix. Then, we prepared flame-retardant thermoplastic corrugated sandwich panels with corresponding flame retardant addition amounts. The limiting oxygen index test, vertical combustion test, cone calorimetry test, and mechanical property test were carried out on each group of samples and control group samples. The results showed that when the flame retardant was added at 25%, the flame retardant level could reach the V0 level. Compared with the control group, the flexural strength and flexural modulus decreased by 2.6%, 14.1%, and 19.9% and 7.3%, 16.1%, and 21.9%, respectively. When the amount of flame retardant was 15%, 20%, and 25%, respectively, the total heat release decreased by 16.3%, 23.5%, and 34.1%, and the maximum heat release rate decreased by 12.5%, 32.4%, and 37.4%, respectively.

Study on the Thermal Stability of the Sweet-Tasting Protein Brazzein Based on Its Structure-Sweetness Relationship.

Brazzein (Brz) is a sweet-tasting protein composed of 54 amino acids and is considered as a potential sugar substitute. The current methods for obtaining brazzein are complicated, and limited information is available regarding its thermal stability. In this study, we successfully expressed recombinant brazzein, achieving a sweetness threshold of 15.2 µg/mL. Subsequently, we conducted heat treatments at temperatures of 80, 90, 95, and 100 °C for a duration of 2 h to investigate the structural changes in the protein. Furthermore, we employed hydrogen-deuterium exchange coupled to mass spectrometry (HDX-MS) to analyze the effect of heating on the protein structure-sweetness relationships. Our results indicated that the thermal inactivation process primarily affects residues 6-14 and 36-45 of brazzein, especially key residues Tyr8, Tyr11, Ser14, Glu36, and Arg43, which are closely associated with its sweetness. These findings have significant implications for improving the thermal stability of brazzein.

Aeration-driven piezoelectric activation of peroxymonosulfate achieves effective mitigation of antibiotic resistance dissemination.

The antibiotic resistance dissemination in water has become a globally concerned issue, and the wastewater discharge, especially medical wastewater, is considered as one of the most important sources for antibiotic resistance genes (ARGs). However, the effectiveness of current disinfection techniques in the ARGs reduction still remains controversial. In this study, a novel aeration-driven piezoelectric peroxymonosulfate (PMS) activation system using oxygen-vacancy engineered BaTiO3 (BTO) was developed to effectively eliminate antibiotic resistant bacteria (ARB) and ARGs from water. The ARB can be completely inactivated and ∼3.0 logs of ARGs can be removed by the PMS/BTO/aeration system within 1 h, and the spent BTO nanoparticles can be facilely reused after simple rinsing. The aeration can not only provide the driving force for the piezocatalytic process but also more dissolved oxygen in water that played an important role in the generation of free radicals. The radical quenching experiments and electron spin-resonance (ESR) confirmed that all the free radicals, including singlet oxygen (1O2), hydroxyl radical (OH•), sulfate radical (SO4•-) and superoxide radical (•O2-), contributed to the ARGs reduction and 1O2 radicals were identified as the dominant active species. This work provides a high-efficiency and energy saving approach for the mitigation of ARGs from water as the universal use of aeration in water treatment processes and the good reusability of BTO nanoparticles.

Mitochondria-lysosome-extracellular vesicles axis and nanotheranostics in neurodegenerative diseases.

The intricate functional interactions between mitochondria and lysosomes play a pivotal role in maintaining cellular homeostasis and proper cellular functions. This dynamic interplay involves the exchange of molecules and signaling, impacting cellular metabolism, mitophagy, organellar dynamics, and cellular responses to stress. Dysregulation of these processes has been implicated in various neurodegenerative diseases. Additionally, mitochondrial-lysosomal crosstalk regulates the exosome release in neurons and glial cells. Under stress conditions, neurons and glial cells exhibit mitochondrial dysfunction and a fragmented network, which further leads to lysosomal dysfunction, thereby inhibiting autophagic flux and enhancing exosome release. This comprehensive review synthesizes current knowledge on mitochondrial regulation of cell death, organelle dynamics, and vesicle trafficking, emphasizing their significant contributions to neurodegenerative diseases. Furthermore, we explore the emerging field of nanomedicine in the management of neurodegenerative diseases. The review provides readers with an insightful overview of nano strategies that are currently advancing the mitochondrial-lysosome-extracellular vesicle axis as a therapeutic approach for mitigating neurodegenerative diseases.

Transcriptomic and metabolomic analysis reveals the influence of carbohydrates on lignin degradation mediated by Bacillus amyloliquefaciens.

Introduction: Ligninolytic bacteria can secrete extracellular enzymes to depolymerize lignin into small-molecular aromatics that are subsequently metabolized and funneled into the TCA cycle. Carbohydrates, which are the preferred carbon sources of bacteria, influence the metabolism of lignin-derived aromatics through bacteria. Methods: In this study, untargeted metabolomics and transcriptomics analyses were performed to investigate the effect of carbohydrates on lignin degradation mediated by Bacillus amyloliquefaciens MN-13, a strain with lignin-degrading activity that was isolated in our previous work. Results: The results demonstrated that the cell growth of the MN-13 strain and lignin removal were promoted when carbohydrates such as glucose and sodium carboxymethyl cellulose were added to an alkaline lignin-minimal salt medium (AL-MSM) culture. Metabolomics analysis showed that lignin depolymerization took place outside the cells, and the addition of glucose regulated the uptake and metabolism of lignin-derived monomers and activated the downstream metabolism process in cells. In the transcriptomics analysis, 299 DEGs were screened after 24 h of inoculation in AL-MSM with free glucose and 2 g/L glucose, respectively, accounting for 8.3% of the total amount of annotated genes. These DEGs were primarily assigned to 30 subcategories, including flagellar assembly, the PTS system, RNA degradation, glycolysis/gluconeogenesis, the TCA cycle, pyruvate metabolism, and tryptophan metabolism. These subcategories were closely associated with the cell structure, generation of cellular energy, and precursors for biosynthetic pathways, based on a - log 10 (P adjust) value in the KEGG pathway analysis. Conclusion: In summary, the addition of glucose increased lignin degradation mediated by the MN-13 strain through regulating glycolysis, TCA cycle, and central carbon metabolism.

Improved lipase performance by covalent immobilization of Candida antarctica lipase B on amino acid modified microcrystalline cellulose as green renewable support.

Development of immobilized lipase with excellent catalytic performance and low cost is the major challenge for large-scale industrial applications. In this study, green renewable microcrystalline cellulose (MCC) that was hydrophobically modified with D-alanine (Ala) or L-lysine (Lys) was used for immobilizing Candida antarctica lipase B (CALB). The improved catalytic properties were investigated by experimental and computational methods. CALB immobilized on MCC-Ala with higher hydrophobicity showed better catalytic activity than CALB@MCC-Lys because the increased flexibility of the lid region of CALB@MCC-Ala favored the formation of open conformation. Additionally, the low root mean square deviation and the high ß-sheet and α-helix contents of CALB@MCC-Ala indicated that the structure became more stable, leading to a significantly enhanced stability (54.80% and 90.90% relative activity at 70 °C and pH 9.0, respectively) and good reusability (48.92% activity after 5 cycles). This study provides a promising avenue to develop immobilized lipase with high catalytic properties for industry applications.

CsPAT1, a GRAS transcription factor, promotes lignin accumulation by antagonistic interacting with CsWRKY13 in tea plants.

Lignin is an important component of plant cell walls and plays crucial roles in the essential agronomic traits of tea quality and tenderness. However, the molecular mechanisms underlying the regulation of lignin biosynthesis in tea plants remain unclear. CsWRKY13 acts as a negative regulator of lignin biosynthesis in tea plants. In this study, we identified a GRAS transcription factor, phytochrome A signal transduction 1 (CsPAT1), that interacts with CsWRKY13. Silencing CsPAT1 expression in tea plants and heterologous overexpression in Arabidopsis demonstrated that CsPAT1 positively regulates lignin accumulation. Further investigation revealed that CsWRKY13 directly binds to the promoters of CsPAL and CsC4H and suppresses transcription of CsPAL and CsC4H. CsPAT1 indirectly affects the promoter activities of CsPAL and CsC4H by interacting with CsWRKY13, thereby facilitating lignin biosynthesis in tea plants. Compared with the expression of CsWRKY13 alone, the co-expression of CsPAT1 and CsWRKY13 in Oryza sativa significantly increased lignin biosynthesis. Conversely, compared with the expression of CsPAT1 alone, the co-expression of CsPAT1 and CsWRKY13 in O. sativa significantly reduced lignin accumulation. These results demonstrated the antagonistic regulation of the lignin biosynthesis pathway by CsPAT1 and CsWRKY13. These findings improve our understanding of lignin biosynthesis mechanisms in tea plants and provide insights into the role of the GRAS transcription factor family in lignin accumulation.

C1ql3 knockout affects microglia activation, neuronal integrity, and spontaneous behavior in Wistar rats.

BACKGROUND: C1QL3 is widely expressed in the brain and is specifically produced by a subset of excitatory neurons. However, its function is still not clear. We established C1ql3-deficient rats to investigate the role of C1QL3 in the brain. METHODS: C1ql3 knockout (KO) rats were generated using CRISPR/Cas9. C1ql3 KO was determined by polymerase chain reaction (PCR), DNA sequencing, and western blotting. Microglia morphology and cytokine expression with or without lipopolysaccharide (LPS) stimulus were analyzed using immunohistochemistry and real-time PCR. The brain structure changes in KO rats were examined using magnetic resonance imaging. Neuronal architecture alteration was analyzed by performing Golgi staining. Behavior was evaluated using the open field test, Morris water maze test, and Y maze test. RESULTS: C1ql3 KO significantly increased the number of ramified microglia and decreased the number of hypertrophic microglia, whereas C1ql3 KO did not influence the expression of pro-inflammatory factors and anti-inflammatory factors except IL-10. C1ql3 KO brains had more amoeboid microglia types and higher Arg-1 expression compared with the WT rats after LPS stimulation. The brain weights and HPC sizes of C1ql3 KO rats did not differ from WT rats. C1ql3 KO damaged neuronal integrity including neuron dendritic arbors and spine density. C1ql3 KO rats demonstrated an increase in spontaneous activity and an impairment in short working memory. CONCLUSIONS: C1ql3 KO not only interrupts the neuronal integrity but also affects the microglial activation, resulting in hyperactive behavior and impaired short memory in rats, which highlights the role of C1QL3 in the regulation of structure and function of both neuronal and microglial cells.