Proteolysis in plant immunity.

Compared with transcription and translation, protein degradation machineries can act faster and be targeted to different subcellular compartments, enabling immediate regulation of signaling events. It is therefore not surprising that proteolysis has been used extensively to control homeostasis of key regulators in different biological processes and pathways. Over the past decades, numerous studies have shown that proteolysis, where proteins are broken down to peptides or amino acids through ubiquitin-mediated degradation systems and proteases, is a key regulatory mechanism to control plant immunity output. Here, we briefly summarize the roles various proteases play during defense activation, focusing on recent findings. We also update the latest progress of ubiquitin-mediated degradation systems in modulating immunity by targeting plant membrane-localized pattern recognition receptors (PRRs), intracellular nucleotide-binding domain leucine-rich repeat receptors (NLRs), and downstream signaling components. Additionally, we highlight recent studies showcasing the importance of proteolysis in maintaining broad-spectrum resistance without obvious yield reduction, opening new directions for engineering elite crops that are resistant to a wide range of pathogens with high yield.

Dexmedetomidine inhibited arrhythmia susceptibility to adrenergic stress in RyR2R2474S mice through regulating the coupling of membrane potential and intracellular calcium.

BACKGROUND: Dexmedetomidine (DEX), a highly selective α2-adrenoceptor agonist, can decrease the incidence of arrhythmias, such as catecholaminergic polymorphic ventricular tachycardia (CPVT). However, the underlying mechanisms by which DEX affects cardiac electrophysiological function remain unclear. METHODS: Ryanodine receptor (RyR2) heterozygous R2474S mice were used as a model for CPVT. WT and RyR2R2474S/+ mice were treated with isoproterenol (ISO) and DEX, and electrocardiograms were continuously monitored during both in vivo and ex vivo experiments. Dual-dye optical mapping was used to explore the anti-arrhythmic mechanism of DEX. RESULTS: DEX significantly reduced the occurrence and duration of ISO-induced of VT/VF in RyR2R2474S/+ mice in vivo and ex vivo. DEX remarkably prolonged action potential duration (APD80) and calcium transient duration (CaTD80) in both RyR2R2474S/+ and WT hearts, whereas it reduced APD heterogeneity and CaT alternans in RyR2R2474S/+ hearts. DEX inhibited ectopy and reentry formation, and stabilized voltage-calcium latency. CONCLUSION: DEX exhibited an antiarrhythmic effect through stabilizing membrane voltage and intracellular Ca2+. DEX can be used as a beneficial perioperative anesthetic for patients with CPVT or other tachy-arrhythmias.

scRNA-Seq: First Atlas and Cellular Landscape of Lacrimal Sac: Implications in Primary Acquired Nasolacrimal Duct Obstruction Pathogenesis.

Purpose: The aim of this study was to describe the transcriptional changes of individual cellular components in the lacrimal sac in patients with primary acquired nasolacrimal duct obstruction (PANDO) and attempt to construct the first lacrimal sac cellular atlas to elucidate the potential mechanisms that may drive the disease pathogenesis. Methods: Lacrimal sac samples were obtained intra-operatively during the endoscopic dacryocystorhinostomy (EnDCR) procedure from five patients. Single-cell RNA sequencing was performed to analyze each individual cell population including epithelial and immune cells during the early inflammatory and late inflammatory phases of the disease. Results: Eleven cell types were identified among 25,791 cells. T cells and B cells were the cell populations with the greatest variation in cell numbers between the two phases and were involved in immune response and epithelium migration-related pathways. The present study showed that epithelial cells highly expressed the genes of senescence-associated secretory phenotype (SASP) and were involved in influencing the inflammation, neutrophil chemotaxis, and migration during the late inflammatory stage. Enhanced activity of CXCLs-CXCRs between the epithelial cells and neutrophils was noted by the cell-cell communication analysis and is suspected to play a role in inflammation by recruiting more neutrophils. Conclusions: The study presents a comprehensive single-cell landscape of the lacrimal sac cells in different phases of PANDO. The contribution of T cells, B cells, and epithelial cells to the inflammatory response, and construction of the intercellular signaling networks between the cells within the lacrimal sac has further enhanced the present understanding of the PANDO pathogenesis.

Primary Lacrimal Sac Tumors with Extension into Vicinity: Outcomes with Endoscopy-Assisted Modified Weber-Ferguson's Approach.

PURPOSE: To evaluate the outcomes of endoscopy-assisted modified Weber-Ferguson's approach in the management of primary lacrimal sac tumors with extension into the neighboring tissues. METHODS: A retrospective interventional study was performed on all patients with lacrimal sac tumors treated with the endoscopy-assisted modified Weber-Ferguson approach between January 2010 and June 2022 at the Shanghai Ninth People's Hospital, China. Data assessed include demographics, clinical presentations, imaging features, surgical techniques, histopathology, adjuvant modalities of management, complications, and outcomes. RESULTS: A total of 13 patients were included in the analysis. Epiphora and palpable mass lesion were the presenting complaint in 84.6% (11/13) of the patients. Nearly half of the patients (46.1%, 6/13) were misdiagnosed as lacrimal duct obstruction. All the lacrimal sac tumors in the present series showed uneven enhancement on T1-weighted MRI imaging. Postoperatively, 84.6% (11/13) patients recovered well with excellent esthetics and were disease-free after a mean follow-up of 58.6 months. Two patients who underwent additional exenteration developed recurrence and succumbed (at 41 and 96 months follow up) while they were on palliative chemoradiation. CONCLUSION: The endoscopic-assisted modified Weber-Fergusson surgical approach is effective in providing better visibility and accessibility to lacrimal sac tumors with extension into neighboring tissue.

Evaluation of primary acquired nasolacrimal duct obstruction: Comparison of CT-DCG and dacryoendoscopy in accurately localizing the lacrimal drainage obstructions.

OBJECTIVE: To correlate and evaluate the power and limitations of CT-DCG in determining the level and type of lacrimal duct obstruction in comparison to dacryoendoscopy in patients clinically suspected to be having partial or complete primary acquired nasolacrimal duct obstruction (PANDO). METHODS: A retrospective chart review was performed on 1232 lacrimal drainage systems of 957 patients who suffered from primary acquired nasolacrimal duct obstruction (PANDO) at Shanghai Ninth People's Hospital. Patients were examined with CT-DCG and correlated with dacryoendoscopy and the findings of clinical examination. RESULTS: Of the studied patients, 173 were men and 784 were women with an age range of 18-93 years. Of the 1232 lacrimal pathways, good CT-DCG images could be obtained in 980 cases and dacryoendoscopy in 957 cases. Of these complete obstructions were noted in 81% (794/980), and partial obstructions were identified in 19% (186/980) with CT-DCG. CT-DCG and dacryoendoscopy showed 68.4% agreement for the type of the obstruction and 63% for the level of the obstruction. The majority of the obstructions occurred at the sac-duct junction (62.5%) followed by the upper half of the nasolacrimal duct (27.5%). There was a significant difference in the correlation of the obstruction type with age group and with the duration of symptoms. As the duration of symptoms increased, the proportion of complete lacrimal duct obstructions as shown on CT-DCG images increased and the proportion of incomplete obstruction decreased (p = 0.015). CONCLUSIONS: The junction of lacrimal sac and nasolacrimal duct was the most common obstruction site. Age and the duration of symptoms influenced the type of obstruction noted. The degree and level of agreement between the investigations was moderate. A combination of CT-DCG and Dacryoendoscopy could together identify the location more accurately.

Spinal interleukin-16 mediates inflammatory pain via promoting glial activation.

Proinflammatory cytokines are crucial contributors to neuroinflammation in the development of chronic pain. Here, we identified il16, which encodes interleukin-16 (IL-16), as a differentially expressed gene in spinal dorsal horn of a complete Freund's Adjuvant (CFA) inflammatory pain model in mice by RNA sequencing. We further investigated whether and how IL-16 regulates pain transmission in the spinal cord and contributes to the development of inflammatory pain hypersensitivity. RNA sequencing and bioinformatics analysis revealed elevated IL-16 transcript levels in the spinal dorsal horn after CFA injection. This increase was further confirmed by qPCR, immunofluorescence, and western blotting. Knockdown of IL-16 by intrathecal injection of IL-16 siRNA not only attenuated CFA-induced mechanical and thermal pain hypersensitivity, but also inhibited enhanced c-fos expression and glial activation in the spinal dorsal horn in male mice injected with CFA. Moreover, exogenous IL-16 induced nociceptive responses and increased c-fos expression and glial activation in spinal dorsal horn. This effect was largely impaired when CD4, the binding receptor for IL-16, was inhibited. In addition, CD4 expression was upregulated in the spinal dorsal horn after CFA injection and CD4 was present in microglia and in contact with astrocytes and activated spinal neurons. Taken together, these results suggest that enhanced IL-16-CD4 signaling triggers pain and activates microglia and astrocytes in the spinal dorsal horn, thus contributing to inflammatory pain. IL-16 may serve as a promising target for the treatment of inflammatory pain.

Immune receptor mimicking hormone receptors: a new guarding strategy.

Plant intracellular nucleotide-binding domain leucine-rich repeat (NLR) receptors play crucial roles in immune responses against pathogens. How diverse NLRs recognize different pathogen effectors remains a significant question. A recent study published in Nature uncovered how pepper NLR Tsw detects phytohormone receptors' interference caused by tomato spotted wilt virus (TSWV) effector, triggering a robust immune response, showcasing a new manner of NLR guarding.

Synthetic exploration of electrophilic xanthylation via powerful N-xanthylphthalimides.

Organic xanthates are broadly applied as synthetic intermediates and bioactive molecules in synthetic chemistry. Electrophilic xanthylation represents a promising approach but has rarely been explored mainly due to the lack of powerful electrophilic reagents. Herein, synthetic exploration of electrophilic xanthylation via powerful N-xanthylphthalimides was investigated. This strategy might provide a new avenue to less-concerned but meaningful electrophilic xanthylation in organic synthesis. With the help of these powerful reagents, electrophilic xanthylation of a wide range of substrates including aryl/alkenyl boronic acids, ß-keto esters, 2-oxindole, and alkyl amines, as well as previously inaccessible phenols (first report) was achieved under mild reaction conditions. Notably, this simple electrophilic xanthylation of alkyl amine substrates will occur in the desulfuration reaction, consistent with the previously reported methods. Similarly, xanthamide and thioxanthate groups could also be transformed into desired nucleophiles via this electrophilic reagent strategy. The broad substrate scope, excellent functional group compatibility and late-stage functionalization of bioactive or functional molecules made them very attractive as general reagents which will allow rapid incorporation of SC(S)R (R = OEt, Oalkyl, NEt2 and SEt) into the target molecules.

SEPSIS LEADS TO IMPAIRED MITOCHONDRIAL CALCIUM UPTAKE AND SKELETAL MUSCLE WEAKNESS BY REDUCING THE MICU1:MCU PROTEIN RATIO.

ABSTRACT: Purpose: Intensive care unit-acquired weakness (ICUAW) is a severe neuromuscular complication that frequently occurs in patients with sepsis. The precise molecular pathophysiology of mitochondrial calcium uptake 1 (MICU1) and mitochondrial calcium uniporter (MCU) in ICUAW has not been fully elucidated. Here, we speculate that ICUAW is associated with MICU1:MCU protein ratio-mediated mitochondrial calcium ([Ca 2+ ] m ) uptake dysfunction. Methods: Cecal ligation and perforation (CLP) was performed on C57BL/6J mice to induce sepsis. Sham-operated animals were used as controls. Lipopolysaccharide (LPS) (5 µg/mL) was used to induce inflammation in differentiated C2C12 myoblasts. Compound muscle action potential (CMAP) was detected using a biological signal acquisition system. Grip strength was measured using a grip-strength meter. Skeletal muscle inflammatory factors were detected using ELISA kits. The cross-sectional area (CSA) of the tibialis anterior (TA) muscle was detected by hematoxylin and eosin staining. Cytosolic calcium ([Ca 2+ ] c ) levels were measured using Fluo-4 AM. Adeno-associated virus (AAV) was injected into TA muscles for 4 weeks to overexpress MICU1 prophylactically. A lentivirus was used to infect C2C12 cells to increase MICU1 expression prophylactically. Findings: The results suggest that sepsis induces [Ca 2+ ] m uptake disorder by reducing the MICU1:MCU protein ratio, resulting in skeletal muscle weakness and muscle fiber atrophy. However, MICU1 prophylactic overexpression reversed these effects by increasing the MICU1:MCU protein ratio. Conclusions: ICUAW is associated with impaired [Ca 2+ ] m uptake caused by a decreased MICU1:MCU protein ratio. MICU1 overexpression improves sepsis-induced skeletal muscle weakness and atrophy by ameliorating the [Ca 2+ ] m uptake disorder.

Differential effects of social isolation on oligodendrocyte development in different brain regions: insights from a canine model.

Social isolation (SI) exerts diverse adverse effects on brain structure and function in humans. To gain an insight into the mechanisms underlying these effects, we conducted a systematic analysis of multiple brain regions from socially isolated and group-housed dogs, whose brain and behavior are similar to humans. Our transcriptomic analysis revealed reduced expression of myelin-related genes specifically in the white matter of prefrontal cortex (PFC) after SI during the juvenile stage. Despite these gene expression changes, myelin fiber organization in PFC remained unchanged. Surprisingly, we observed more mature oligodendrocytes and thicker myelin bundles in the somatosensory parietal cortex in socially isolated dogs, which may be linked to an increased expression of ADORA2A, a gene known to promote oligodendrocyte maturation. Additionally, we found a reduced expression of blood-brain barrier (BBB) structural components Aquaporin-4, Occludin, and Claudin1 in both PFC and parietal cortices, indicating BBB disruption after SI. In agreement with BBB disruption, myelin-related sphingolipids were increased in cerebrospinal fluid in the socially isolated group. These unexpected findings show that SI induces distinct alterations in oligodendrocyte development and shared disruption in BBB integrity in different cortices, demonstrating the value of dogs as a complementary animal model to uncover molecular mechanisms underlying SI-induced brain dysfunction.

Long noncoding RNA antisense noncoding RNA in the INK4 locus inhibition alleviates airway remodeling in asthma through the regulation of the microRNA-7-5p/early growth response factor 3 axis.

Asthma, a chronic inflammatory disease of the airways, clinically manifests as airway remodeling. The purpose of this study was to probe the potential role of long noncoding RNA (lncRNA) antisense noncoding RNA in the INK4 locus (lncRNA ANRIL) in the proliferation and migration of airway smooth muscle cell (ASMC) and to explore its potential mechanisms in asthma. Serum samples were obtained from 30 healthy volunteers and 30 patients with asthma. Additionally, platelet-derived growth factor-BB (PDGF-BB) was used to induce airway remodeling in ASMCs. The level of lncRNA ANRIL and microRNA (miR)-7-5p in serum samples were measured by quantitative reverse transcriptase polymerase chain reaction (qRT-PCR). TargetScan predicted the binding site of miR-7-5p to early growth response factor 3 (EGR3) and validated the results using a dual-luciferase reporter assay. 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-2H-tetrazolium bromide (MTT) and Transwell assays were used to detect cellular proliferation and migration, respectively. Subsequently, changes in proliferation- and migration-related genes were verified using western blot analysis and qRT-PCR. These results indicate that lncRNA ANRIL was upregulated in the serum and PDGF-BB-induced ASMCs of patients with asthma, whereas miR-7-5p expression was reduced. EGR3 was a direct target of miR-7-5p. LncRNA ANRIL silencing inhibited the proliferation or migration of ASMCs induced by PDGF-BB through miR-7-5p upregulation. Mechanistic studies indicated that miR-7-5p inhibits the proliferation or migration of PDGF-BB-induced ASMCs by decreasing EGR3 expression. EGR3 upregulation reverses the role of miR-7-5p in airway remodeling. Thus, downregulation of lncRNA ANRIL inhibits airway remodeling through inhibiting the proliferation and migration of PDGF-BB-induced ASMCs by regulating miR-7-5p/EGR3 signaling.

Auxiliary Carboxylate-Induced Assembly of Calix[6]arene-Polyoxotitanate Hybrid Systems with Photocatalytic Activity in the Oxidation of Sulfides.

This study used the tert-butylcalix[6]arene (TBC[6]) as the ligand and successfully synthesized six TBC[6]-stabilized titanium-oxo clusters (TOCs) by the one-step solvothermal reaction. These six compounds were [Ti4O2(TBC[6])2] (Ti4), {Ti2(TBC[6])(EtO)2(SaH2)2} (Ti2-SA, H2Sa = squaric acid), {Ti2(TBC[6])2(EtO)2(Oa)} (Ti2-OA, H2Oa = oxalic acid), [H2Ti4(TBC[6])(BA)2(EtO)10] (Ti4-BA, HBA = benzoic acid), [Ti6O2(TBC[6])(BA)4(OiPr)10] (Ti6-BA), and [Ti8(TBC[6])2(Sal)4(EtO)16] (Ti8-Sal, H2Sal = salicylic acid). These clusters contain one or two TBC[6] ligands, with the biconical or monoconical configuration, greatly increasing the variety of TOCs it could support. The introduction of auxiliary carboxylic ligands can further stimulate the growth of structures, with the cluster core gradually increased from {Ti-TBC[6]-Ti} to {Ti2-TBC[6]-Ti2}, to {Ti3-TBC[6]-Ti3}, and finally to {Ti3-TBC[6]-Ti2-TBC[6]-Ti3} with 3.1 nm length. Structural regulation may affect their solution stability, absorption spectra, and photocurrent response. The study of catalytic activities shows that these clusters can be used as recyclable heterogeneous photocatalysts for the oxidation of sulfide to sulfoxide. The catalytic efficiency of the TBC[6]-Tix system is closely related to the cluster structure, and the exposure of the Ti site on the catalyst surface can significantly enhance the catalytic activity of the clusters.

Prognostic factors of traumatic optic neuropathy based on multimodal analysis-Especially the influence of postoperative dressing change and optic nerve blood supply on prognosis.

Objective: To investigate the critical prognostic factors of patients with traumatic optic neuropathy (TON) treated with endoscopic transnasal optic canal decompression (ETOCD) and to perform multimodal analysis based on imaging examinations of optical coherence tomography angiography (OCTA) and CT scan. Subsequently, a new prediction model was established. Methods: The clinical data of 76 patients with TON who underwent decompression surgery with the endoscope-navigation system in the Department of Ophthalmology, Shanghai Ninth People's Hospital from January 2018 to December 2021 were retrospectively analyzed. The clinical data included demographic characteristics, reasons for injury, interval between injury and surgery, multimode imaging information of CT scan and OCTA, including orbital fracture, optical canal fractures, vessel density of optic disc and macula, and the times of postoperative dressing change. Binary logistic regression was used to establish a model for best corrected visual acuity (BCVA) after treatment as a predictor of TON outcome. Results: Postoperative BCVA improved in 60.5% (46/76) patients and did not improve in 39.5% (30/76) patients. The times of postoperative dressing change had a significant impact on the prognosis. Other factors affecting the prognosis were microvessel density of the central optic disc, the cause of injury, and the microvessel density above the macula. The area under the raw current curves of the predictive model was 0.7596. Conclusions: The times of dressing changes after the operation, i.e., continuous treatment, is the key factor affecting prognosis. The microvessel density in the center of the optic disc and superior macula, quantitatively analyzed by OCTA, is the prognostic factor of TON and may be used as a prognostic marker of TON.

Targeted OUM1/PTPRZ1 silencing and synergetic CDT/enhanced chemical therapy toward uveal melanoma based on a dual-modal imaging-guided manganese metal-organic framework nanoparticles.

Metastasis and chemical resistance are the most serious problems in the treatment of highly aggressive uveal melanoma (UM). The newly identified lncRNA OUM1 is overexpressed in UM, functions as a catalyst and regulates protein tyrosine phosphatase (PTP) activity by binding to PTP receptor type Z1 (PTPRZ1), which plays an important role in cell proliferation, metastasis and chemotherapy resistance in the UM microenvironment. Hence, siRNAs that selectively knocking down the lncRNA OUM1 (siOUM1) and its target gene PTPRZ1 (siPTPRZ1) were designed to inhibit the OUM1/PTPRZ1 pathway to reduce PTP activity, and this reduction in activity interrupts protein tyrosine phosphorylation, suppresses UM proliferation and metastasis and improves cisplatin sensitivity in UM cells. Then, to overcome the limitations of the difficulty of drug administration and traditional therapeutics, the indocyanine green (ICG)-labeled manganese metal-organic framework (MOF) nanoparticles (NPs) were fabricated and linked with arginine-glycine-aspartate (RGD) peptide to carry siOUM1/siPTPRZ1 and cisplatin to achieve targeted siRNA interference-mediated therapy, enhanced cisplatin therapy and chemodynamic therapy. This NP system also has a dual-modal imaging ability because ICG is a near-infrared region fluorescent dye and manganese has the potential to be used in magnetic resonance imaging. This study verifies the significance of the newly discovered lncRNA OUM1 as a new therapeutic target for aggressive UM and provides a drug delivery NP system for precise treatment of UM accompanied with a dual-modal imaging ability.

A microbial quantity monitoring model based on 3D fluorescence data of the cucumber storeroom gas and its use in providing auxiliary early spoilage warning.

A real-time model for monitoring the microbial quantity based on the microbial intrinsic fluorescence information of cucumber storeroom gas was established. Firstly, 3D fluorescence data of the storeroom gas were collected on different storage days. Secondly, the number of components of a parallel factor model was determined to be 3 using the core consistency diagnostic. Thirdly, parallel factor analysis was used to decompose the fluorescence data to obtain the excitation spectra, emission spectra and concentration scores of 3 components. The positions of the fluorescence peaks were consistent with the fingerprints of tryptophan-like, tyrosine-like and phenylalanine-like substances in the characteristic spectrum of each component. And then the prediction model was constructed by fitting the concentration scores of the 3 components with the microbial quantity, and the coefficient of determination was 98.27%, and the cross-validation determination coefficient could reach 91.97%. Finally, after integrating the predicted value of the microbial quantity and the total chromatism of the cucumber pericarp during cucumber storage, the spoilage date was determined to be the 7th day by K-means clustering. The results show that the monitoring model constructed through distinguishing the fluorescence data of airborne microorganisms can effectively monitor the spoilage process.

Less is More: A Shortcut for Anionocages Design Based on (RPO3 2- )-Monourea Coordination.

Anionocages have been developed as a unique family of hydrogen bonded cages. However, strategies for constructing anionocages are mainly limited to that based on (PO4 3- )-bisurea coordination, neither the ligands nor the anions lack the simplicity and diversity of the maturely developed analogues based on metal coordination (i.e. metallocage). We report herein a more simple strategy for anionocages design based on (RPO3 2- )-monourea coordination, utilizing monourea rather than bisurea as the hydrogen binding donor, and RPO3 2- rather than PO4 3- as the acceptor. Two fluorescent, quadruple helicate anionocages were constructed by a bis-monourea ligand, and dianions PhOPO3 2- (H1 ) or HOPO3 2- (H1A ), respectively, which were capable of encapsulating a series of cation guests. As revealed by molecular modeling, H1 features remarkable guest-adaptive cavity breathing without change of the quadruple helicate topology, which allowed the encapsulation of different sized guests in an "induced fit" manner.

Tidal Breathing Pulmonary Function of Children With Allergic Rhinitis.

In order to investigate the characteristics of tidal breathing pulmonary function in children with allergic rhinitis, and explore its role in the relationship between allergic rhinitis and asthma, we conducted this prospective study from January 4, 2016 to January 30, 2019 in Wuhan children's hospital. In this study, 49 children with simple allergic rhinitis were enrolled in the AR group; 50 children with allergic rhinitis concomitant with asthma were enrolled in the AR&A group; 43 healthy children were recruited in the control group. For individuals in each group, the assessment of tidal breath pulmonary function was performed after enrollment. Then participants in the AR group and control group were followed up for 1 year to observe their frequency of wheezing attacks. The parameters of tI/tE, tPTEF/tE, and VPTEF/VE of AR group were significantly higher than AR&A group (P < 0.001). The reduced proportion of tPTEF/tE and VPTEF/VE. in AR group were higher than that in control group (30.61% vs. 11.63%, P < 0.001; 24.49% vs. 11.63%, P < 0.001, respectively). The proportion of patients with reduced tPTEF/tE and VPTEF/VE who occurred recurrent wheezing was higher than that of patients with normal pulmonary function in AR group(P = 0.008). In conclusion, some children with allergic rhinitis has impaired tidal breathing pulmonary function. Tidal breathing pulmonary function test plays an important role in the diagnosis and assessment of children's airway allergic diseases (AR and asthma).