A calcium-dependent protein kinase regulates the defense response in Citrus sinensis.

Citrus Huanglongbing (HLB), which is caused by Candidatus Liberibacter asiaticus (CLas), is one of the most destructive citrus diseases worldwide, and defense-related Citrus sinensis gene resources remain largely unexplored. Calcium signaling plays an important role in diverse biological processes. In plants, a few calcium-dependent protein kinases (CDPKs/CPKs) have been shown to contribute to defense against pathogenic microbes. The genome of Citrus sinensis encodes dozens of CPKs. In this study, the role of CsCPKs in Citrus sinensis defense was investigated. Silencing of CsCPK6 compromised the induction of defense-related genes in Citrus sinensis. Expression of a constitutively active form of CsCPK6 (CsCPK6CA) triggers the activation of defense-related genes in Citrus sinensis. Complementation of CsCPK6 rescued the defense-related gene induction in an Arabidopsis thaliana cpk4/11 mutant, indicating that CsCPK6 carries CPK activity and is capable of functioning as a CPK in Arabidopsis. Moreover, an effector derived from CLas inhibits defense induced by the expression of CsCPK6CA and autophosphorylation of CsCPK6, which suggests the involvement of CsCPK6 and calcium signaling in defense. These results support the positive role of CsCPK6 in Citrus sinensis defense against CLas, and the auto-inhibitory regulation of CsCPK6 provides a potential genome-editing target for improving the defense of Citrus sinensis.

Super-resolution reconstruction of underwater polarized images with a fused attention mechanism.

The polarization imaging technique leverages the disparity between target and background polarization information to mitigate the impact of backward scattered light, thereby enhancing image quality. However, the imaging model of this method exhibits limitations in extracting inter-image features, resulting in less-than-optimal outcomes in turbid underwater environments. In recent years, machine learning methodologies, particularly neural networks, have gained traction. These networks, renowned for their superior fitting capabilities, can effectively extract information from multiple images. The incorporation of an attention mechanism significantly augments the capacity of neural networks to extract inter-image correlation attributes, thereby mitigating the constraints of polarization imaging methods to a certain degree. To enhance the efficacy of polarization imaging in complex underwater environments, this paper introduces a super-resolution network with an integrated attention mechanism, termed as SRGAN-DP. This network is a fusion of an enhanced SRGAN network and the high-performance deep pyramidal split attention (DPSA) module, also proposed in this paper. SRGAN-DP is employed to perform high-resolution reconstruction of the underwater polarimetric image dataset, constructed specifically for this study. A comparative analysis with existing algorithms demonstrates that our proposed algorithm not only produces superior images but also exhibits robust performance in real-world environments.

Adherence to Reporting Guidelines in Major Otolaryngology Publications.

The aim of this study is to determine the adherence rate to reporting guidelines in published otolaryngology research. We performed an evidence-based review of all original clinical research published in 2021 in five otolaryngology journals for adherence to the appropriate guideline for the study type by evaluating whether the corresponding reporting guideline was mentioned in the body of the published manuscript. There were 1140 original research articles included in this study. Most studies were observational, for which the STROBE reporting guidelines are recommended (n = 791, 70.3%). All studies had an average adherence rate of 16.8% (n = 192/1140). The STROBE adherence rate was 4.9%, with JAMA Otolaryngology having the highest proportion of observation studies using the STROBE guidelines (23/49, 46.9%). Reporting guidelines are important tools to use in presenting original research. The use of these guidelines varies in the otolaryngology literature and highlights the ongoing need to support research reproducibility and usefulness.

Janus Membranes with Asymmetric Superwettability for High-Performance and Long-Term On-Demand Oil/Water Emulsion Separation.

Current single-function superwettable materials are typically designed for either oil removal or water removal and are constrained by oil density, limiting their widespread applications. Janus membranes with opposite wettability on their two surfaces have recently emerged and present attractive opportunities for on-demand oil/water emulsion separation. Here, a combination strategy is introduced to prepare a Janus membrane with asymmetric superwettability for switchable oil/water emulsion separation. A mussel-inspired asymmetric interface introduction cooperating with the sequence-confined surface modification not only brings about an asymmetric superwettability Janus interface but also guarantees an outstanding stable interface and remarkable chemical stability surfaces. Specifically, the superhydrophilic surface with underwater superoleophobicity can separate surfactant-stabilized oil-in-water emulsions. Conversely, other surface displays opposite superhydrophobicity and superoleophilicity to treat surfactant-stabilized water-in-oil emulsions. Significantly, this superwettable Janus membrane presents superior long-term on-demand oil/water emulsion separation without obvious flux decline and high recovery ability because of its superwettability and superior stability. Furthermore, the asymmetric superwettability enhances the interfacial floatability at air-water interfaces, enabling the design of advanced interfacial materials. The as-prepared superwettable Janus membrane has established a cooperated separation system, overcoming the monotony of conventional superwettable membranes and expanding the application of these specialized membranes to oily wastewater treatment.

3-Amide-ß-carbolines block the cell cycle by targeting CDK2 and DNA in tumor cells potentially as anti-mitotic agents.

ß-Carboline alkaloids are natural and synthetic products with outstanding antitumor activity. C3 substituted and dimerized ß-carbolines exert excellent antitumor activity. In the present research, 37 ß-carboline derivatives were synthesized and characterized. Their cytotoxicity, cell cycle, apoptosis, and CDK2- and DNA-binding affinity were evaluated. ß-Carboline monomer M3 and dimer D4 showed selective activity and higher cytotoxicity in tumor cells than in normal cells. Structure-activity relationships (SAR) indicated that the amide group at C3 enhanced the antitumor activity. M3 blocked the A549 (IC50 = 1.44 ± 1.10 µM) cell cycle in the S phase and inhibited A549 cell migration, while D4 blocked the HepG2 (IC50 = 2.84 ± 0.73 µM) cell cycle in the G0/G1 phase, both of which ultimately induced apoptosis. Furthermore, associations of M3 and D4 with CDK2 and DNA were proven by network pharmacology analysis, molecular docking, and western blotting. The expression level of CDK2 was downregulated in M3-treated A549 cells and D4-treated HepG2 cells. Moreover, M3 and D4 interact with DNA and CDK2 at sub-micromolar concentrations in endothermic interactions caused by entropy-driven adsorption processes, which means that the favorable entropy change (ΔS > 0) overcomes the unfavorable enthalpy change (ΔH > 0) and drives the spontaneous reaction (ΔG < 0). Overall, these results clarified the antitumor mechanisms of M3 and D4 through disrupting the cell cycle by binding DNA and CDK2, which demonstrated the potential of M3 and D4 as novel antiproliferative drugs targeting mitosis.

[DNA barcode screening, identification of germplasm resources, and analysis of genetic diversity of Anemarrhena asphodeloides].

Anemarrhena asphodeloides is a common medicinal material used in clinical prescriptions and Chinese patent medicine. In this study, the Illumina platform was used to obtain the chloroplast genome sequences of seven kinds of A. asphodeloides from different areas. The specific DNA barcodes were screened by comparative genomics analysis, and the DNA barcodes were used to identify the germplasm resources and analyze the genetic diversity of A. asphodeloides samples from different areas in China. All the seven chloroplast genomes had a ring structure. The total length was 156 801-156 930 bp, and 113 genes were annotated, including 79 protein-coding genes, 30 tRNA genes, and four rRNA genes. The comparative genomics analysis showed that rps16, trnG-GCC, atpF, rpoB, ycf3, rpl16, ndhF, trnS-GCU＿trnG-GCC, petN-psbM, and ndhF-rpl32 were potential candidates for specific DNA barcodes of A. asphodeloides. In this study, the second intron of ycf3 and atpF intron sequences with a sequence length of 700-800 bp and easy amplification were selected for polymerase chain reaction(PCR) amplification and sequencing of 594 samples from 26 areas. The sequence analysis showed that six and eight haplotypes of ycf3 and atpF sequences could be identified, respectively, and 17 haplotypes could be identified by combined analysis of the two sequences, which were named Hap1-Hap17. The haplotype diversity(H_d), nucleotide diversity(P_i), and genetic distance of A. asphodeloides in 26 populations were 0.68, 0.93×10~(-3), and 0-0.003 1, respectively, indicating that the genetic diversity within the species of A. asphodeloides is rich. The intermediary adjacent network analysis showed that Hap5 was the oldest haplotype, which was mainly distributed in Yixian county of Baoding, Hebei province, Hequ county of Xinzhou, Shanxi province, and Xiangfen county of Linfen, Shanxi province. This study has important guiding significance for the identification of A. asphodeloides species, the protection and development of germplasm resources, and the identification of production areas, and it provides a research basis for further revealing the genetic evolution law of A. asphodeloides.

miR-30c-5p Protects from Brain Injury After Cerebral Aneurysmal Subarachnoid Hemorrhage Through the ATG5/ATG12 Pathway.

Objective: To assess the role of miR-30c-5p in subarachnoid hemorrhage (SAH) and its possible mechanism. Methods: We established a SAH model by injecting fresh arterial non-heparinized blood into the anterior cistern of the optic chiasm of healthy Sprague-Dawley rats. Next, we treated the rats with a miR-30c-5p inhibitor or miR-30c-5p mimics. We then assessed behavior, serum lactate dehydrogenase levels, albumin expression, neuronal degeneration, neuronal apoptosis, neuronal survival, and the cerebral edema index in the SAH model rats. We identified downstream target genes of miR-30c-5p using the Targetscan database and confirmed them via luciferase reporter assay. Finally, we assessed the effect of these targeted genes on brain injury in SAH rats through a recovery assay. Results: Our results showed that the overexpression of miR-30c-5p in brain tissue 24h after SAH prevented brain injury, reduced inflammation levels and nerve function scores, inhibited neuronal apoptosis, and improved neuronal survival. Meanwhile, inhibiting miR-30c-5p yielded opposite effects. Two genes related to the autophagy pathway, ATG5 and ATG12, were identified as miR-30c-5p downstream target genes. Silencing ATG5 and ATG12 alleviated brain injury induced by knocking down miR-30c-5p. Conclusion: Our findings suggest that miR-30c-5p protects from SAH-induced brain injury by inhibiting the ATG5/ATG12 pathway and it may serve as a new diagnostic maker or target for treatment of SAH patients.

Economic Evaluation of Pediatric Tracheostomy: A Cost of Illness Analysis.

Objective: This study aimed to determine the direct costs of pediatric tracheostomy care within a health care system. Study Design: Prospective analysis. Setting: Academic children's hospital. Methods: Costs associated with caring for pediatric tracheostomy patients under 18 years were analyzed between 2015 and 2021. Direct costs were calculated using the Medicare/Medicaid charges-to-costs ratio for various visit types. Costs were estimated using generalized linear equations, accounting for confounders. Results: A total of 297 children underwent tracheostomy at a median age of 0.94 years. The median follow-up was 2.5 years, resulting in 13,966 visits (mean = 41). The total cost was $321 million. The initial admission accounted for 72% ($231 million) of costs while other inpatient admissions added 24% ($78 million). Emergency department, observation, and outpatient visits comprised 4% of costs. The length of stay (LOS) was the primary cost driver for inpatient visits. Each additional hospital day increased costs by roughly $1195, and each extra admission added about $130,223 after adjusting for confounders. Respiratory failure and infections were the primary reasons for 67% of subsequent admissions. Conclusion: Pediatric tracheostomy care generated over $300 million in direct costs over 5 years. Inpatient stays constituted 96% of these costs, with the LOS being a major factor. To reduce direct health expenditures for these patients, the focus should be on minimizing admissions.

Inhibition of quorum sensing serves as an effective strategy to mitigate the risks of human bacterial pathogens in soil.

The coexistence of antibiotic resistance genes (ARGs), mobile genetic elements (MGEs), and virulence factor genes (VFGs) in human bacterial pathogens (HBPs) increases their risks to ecological security and human health and no effective strategy is available. Herein, we demonstrated two typical quorum sensing (QS) interfering agents, 4-nitropyridine-N-oxide (4-NPO, a QS inhibitor) and Acylase Ⅰ (a quorum quenching (QQ) enzyme), effectively decreased the abundance of HBPs by 48.30% and 72.54%, respectively, which was accompanied by the reduction of VFGs, ARGs, and MGEs. The decrease in QS signals mediated by QS interfering agents disturbed bacterial communication and inhibited biofilm formation. More importantly, QS interfering agents reduced the intra-species and inter-species conjugation frequencies among bacteria, considerably inhibiting the dissemination of ARGs and VFGs via horizontal gene transfer. Furthermore, the QS interfering agents did not significantly affect the metabolic function of other nonpathogenic microorganisms in the soil. Collectively, our study provides an effective and eco-friendly strategy to mitigate the risks of HBPs in soil.

Target Separation and Potential Anticancer Activity of Withanolide-Based Glucose Transporter Protein 1 Inhibitors from Physalis angulata var. villosa.

The glucose transporter 1 (GLUT1) protein is involved in the basal-level absorption of glucose in tumor cells. Inhibiting GLUT1 decreases tumor cell proliferation and induces tumor cell damage. Natural GLUT1 inhibitors have been studied only to a small extent, and the structures of known natural GLUT1 inhibitors are limited to a few classes of natural products. Therefore, discovering and researching other natural GLUT1 inhibitors with novel scaffolds are essential. Physalis angulata L. var. villosa is a plant known as Mao-Ku-Zhi (MKZ). Withanolides are the main phytochemical components of MKZ. MKZ extracts and the components of MKZ exhibited antitumor activity in recent pharmacological studies. However, the antitumor-active components of MKZ and their molecular mechanisms remain unknown. A cell membrane-biomimetic nanoplatform (CM@Fe3O4/MIL-101) was used for target separation of potential GLUT1 inhibitors from MKZ. A new withanolide, physagulide Y (2), together with six known withanolides (1, 3-7), was identified as a potential GLUT1 inhibitor. Physagulide Y was the most potent GLUT1 inhibitor, and its antitumor activity and possible mechanism of action were explored in MCF-7 human cancer cells. These findings advance the development of technologies for the targeted separation of natural products and identify a new molecular framework for the investigation of natural GLUT1 inhibitors.

A New Porphyrin-based Covalent Organic Framework with High Iodine Capture Capacity and I-doping Enhanced Conductivity.

Covalent organic frameworks (COFs) are porous organic materials with well-defined and uniform structure. The material is an excellent candidate as a solid adsorbent for iodine adsorption. In the present study, we report the synthesis of COF with porphyrin moiety, TF-TA-COF, by solvothermal reaction, which was characterized by XRD, solid-state 13 C NMR, IR, TGA, and nitrogen adsorption-desorption analysis. TF-TA-COF showed a high specific surface area of 443 m2 g-1 , and exhibited good adsorption performance for iodine vapor, with an adsorption capacity of 2.74 g g-1 . XPS and Raman spectrum indicated that a hybrid of physisorption and chemisorption took place between host COF and iodine molecules. The electric properties of iodine-loaded TF-TA-COF were also studied. After doped with iodine, the conductivity of the material increased by more than 5 orders of magnitude. The photoconductivity of I2 -doped COF was also studied and TF-TA-COF showed doping-enhanced photocurrent generation.

Side effects of X-ray irradiation on flight ability of Cydia pomonella moth.

BACKGROUND: The sterile insect technique (SIT) has proven to be an effective approach in managing the population of major invasive pests. Our previous studies showed that irradiation of Cydia pomonella males at a dosage of 366 Gy X-rays resulted in complete sterility. However, the mating competitiveness of sterilized males is significantly compromised, which can be attributed to a decline in their ability to fly. RESULTS: In this study, we examined the flight patterns of both male and female adults of C. pomonella. The results revealed significant variations in the average flight speed of both genders at different stages of maturity, with females displaying longer flight duration and covering greater distances. Effect of irradiation on the flight performance of 3-day-old male moths was further evaluated, as they demonstrated the longest flight distance. The findings indicated a significant decrease in flight distance, duration, and average speed, due to wing deformities caused by irradiation, which also limited the dispersal distance of moths in orchards, as indicated by the mark-and-recapture assay. Reverse-transcription quantitative polymerase chain reaction analysis revealed a down-regulation of flight-related genes such as Flightin, myosin heavy chain, and Distal-less following radiation exposure. CONCLUSION: These findings demonstrate that X-ray irradiation at a radiation dose of 366 Gy has a detrimental effect on the flight ability of male C. pomonella adults. These insights not only contribute to a better understanding of how radiation sterilization diminishes the mating competitiveness of male moths, but also aid in the development and improvement of SIT practices for the effective control of C. pomonella. © 2023 Society of Chemical Industry.

Association of carotid wall shear stress measured by vector flow mapping technique with ba-PWV: a pilot study.

Objective: Arterial stiffness is an important tissue biomarker of the progression of atherosclerotic diseases. Brachial-ankle pulse wave velocity (ba-PWV) is a gold standard of arterial stiffness measurement widely used in Asia. Changes in vascular wall shear stress (WSS) lead to artery wall remodeling, which could give rise to an increase in arterial stiffness. The study aimed to explore the association between ba-PWV and common carotid artery (CCA) WSS measured by a newly invented vascular vector flow mapping (VFM) technique. Methods: We included 94 subjects free of apparent cardiovascular disease (CVD) and divided them into a subclinical atherosclerosis (SA) group (N = 47) and non subclinical atherosclerosis (NSA) group (N = 47). CCA WSS was measured using the VFM technique. Bivariate correlations between CCA WSS and other factors were assessed with Pearson's, Spearman's, or Kendall's coefficient of correlation, as appropriate. Partial correlation analysis was conducted to examine the influence of age and sex. Multiple linear stepwise regression was used for the analysis of independent determinants of CCA WSS. Receiver operating characteristic (ROC) analysis was performed to find the association between CCA WSS and 10-year CVD risk. Results: The overall subjects had a mean age of 47.9 ± 11.2 years, and males accounted for 52.1%. Average systolic CCA WSS was significantly correlated with ba-PWV (r = -0.618, p < 0.001) in the SA group. Multiple linear stepwise regression analysis confirmed that ba-PWV was an independent determinant of average systolic CCA WSS (ß = -0.361, p = 0.003). The area under the curve (AUC) of average systolic CCA WSS for 10-year CVD risk ≥10% was 0.848 (p < 0.001) in the SA group. Conclusions: Average systolic CCA WSS was significantly correlated with ba-PWV and was associated with 10-year CVD risk ≥10% in the SA group. Therefore, CCA WSS measured by the VFM technique could be used for monitoring and screening subjects with potential CVD risks.

Tapered Fiber Bioprobe Based on U-Shaped Fiber Transmission for Immunoassay.

In this paper, a tapered fiber bioprobe based on Mach-Zehnder interference (MZI) is proposed. To retain the highly sensitive straight-tapered fiber MZI sensing structure, we designed a U-shaped transmission fiber structure for the collection of optical sensing signals to achieve a miniature-insert-probe design. The spectrum responses from the conventional straight-tapered fiber MZI sensor and our proposed sensor were compared and analyzed, and experimental results showed that our proposed sensor not only has the same sensing capability as the straight-tapered fiber sensor, but also has the advantages of being flexible, convenient, and less liquid-consuming, which are attributed to the inserted probe design. The tapered fiber bioprobe obtained a sensitivity of 1611.27 nm/RIU in the refractive index detection range of 1.3326-1.3414. Finally, immunoassays for different concentrations of human immunoglobulin G were achieved with the tapered fiber bioprobe through surface functionalization, and the detection limit was 45 ng/mL. Our tapered fiber bioprobe has the insert-probe advantages of simpleness, convenience, and fast operation. Simultaneously, it is low-cost, highly sensitive, and has a low detection limit, which means it has potential applications in immunoassays and early medical diagnosis.

Corrigendum: Identification of hub cuproptosis related genes and immune cell infiltration characteristics in periodontitis.

[This corrects the article DOI: 10.3389/fimmu.2023.1164667.].

A complete MAP kinase cascade controls hyphopodium formation and virulence of Verticillium dahliae.

Phytopathogens develop specialized infection-related structures to penetrate plant cells during infection. Different from phytopathogens that form appressoria or haustoria, the soil-borne root-infecting fungal pathogen Verticillium dahliae forms hyphopodia during infection, which further differentiate into penetration pegs to promote infection. The molecular mechanisms underlying the regulation of hyphopodium formation in V. dahliae remain poorly characterized. Mitogen-activated protein kinases (MAPKs) are highly conserved cytoplasmic kinases that regulate diverse biological processes in eukaryotes. Here we found that deletion of VdKss1, out of the five MAPKs encoded by V. dahliae, significantly impaired V. dahliae hyphopodium formation, in vitro penetration, and pathogenicity in cotton plants. Constitutive activation of MAPK kinase (MAPKK) VdSte7 and MAPK kinase kinase (MAPKKK) VdSte11 specifically activate VdKss1. Deletion of VdSte7 or VdSte11 resulted in a phenotype similar to that of the mutant with VdKss1 deletion. Thus, this study demonstrates that VdSte11-VdSte7-VdKss1 is a core MAPK cascade that regulates hyphopodium formation and pathogenicity in V. dahliae. Supplementary Information: The online version contains supplementary material available at 10.1007/s42994-023-00102-y.

The Effect of X-ray Irradiation on the Fitness and Field Adaptability of the Codling Moth: An Orchard Study in Northeast China.

The codling moth, Cydia pomonella (L.), is an invasive agricultural pest of pome fruits and walnuts in China that threatens the apple industry in the Loess Plateau and Bohai Bay; it has developed resistance to many insecticides. Sterile insect technique (SIT) combined with area-wide integrated pest management (AW-IPM) can reduce the risk of resistance to insecticides and effectively control some insect pest species. Our previous laboratory experiment found that irradiation with 366 Gy of X-ray caused the males of the codling moth to become sterile. However, the sterility and adaptability of males after being irradiated with 366 Gy X-ray in the field are still unclear. In this study, we investigated the effect of X-ray irradiation on the fitness of male adults that emerged from pupae irradiated with 366 Gy to explore their adaptability and mating competitiveness, and to examine the effect of releasing sterile male insects in orchards in northeast China on the fruit infestation rate of the Nanguo pear. The results showed that 366 Gy of X-ray irradiation significantly reduced the mating competitiveness of males and the hatching rate of the eggs laid by females pairing with sterile males. Meanwhile, the lifespan of the sterile male moths was significantly shorter than that of the normal ones in the field. A pilot test showed that the release twice of sterile male moths in the orchards had no significant effect on the fruit infestation rate. Our field experiments provide a scientific basis for the further optimization of the SIT technology program for controlling C. pomonella.

Ultrasensitive and Specific Clustered Regularly Interspaced Short Palindromic Repeats Empowered a Plasmonic Fiber Tip System for Amplification-Free Monkeypox Virus Detection and Genotyping.

The urgent necessity for highly sensitive diagnostic tools has been accentuated by the ongoing mpox (monkeypox) virus pandemic due to the complexity in identifying asymptomatic and presymptomatic carriers. Traditional polymerase chain reaction-based tests, despite their effectiveness, are hampered by limited specificity, expensive and bulky equipment, labor-intensive operations, and time-consuming procedures. In this study, we present a clustered regularly interspaced short palindromic repeats (CRISPR)/Cas12a-based diagnostic platform with a surface plasmon resonance-based fiber tip (CRISPR-SPR-FT) biosensor. The compact CRISPR-SPR-FT biosensor, with a 125 µm diameter, offers high stability and portability, enabling exceptional specificity for mpox diagnosis and precise identification of samples with a fatal mutation site (L108F) in the F8L gene. The CRISPR-SPR-FT system can analyze viral double-stranded DNA from mpox virus without amplification in under 1.5 h with a limit of detection below 5 aM in plasmids and about 59.5 copies/µL when in pseudovirus-spiked blood samples. Our CRISPR-SPR-FT biosensor thus offers fast, sensitive, portable, and accurate target nucleic acid sequence detection.

Highly defective copper-based metal-organic frameworks for the efficient adsorption and detection of organophosphorus pesticides: An experimental and computational investigation.

Organophosphorus pesticide (OP) residues pose a serious threat to human health, motivating the search for novel adsorbents and detection methods. Herein, defective copper-based metal organic frameworks (Cu-MOFs) were synthesized by the reaction of Cu2+ ions and 1,3,5-benzenetricarboxylate linkers in the presence of acetic acid. As the amount of acetic acid increased, the crystallization kinetics and morphology of the Cu-MOFs changed, leading to mesoporous Cu-MOFs with many large surface pores (defects). Adsorption studies of OPs revealed the defective Cu-MOFs showed faster pesticide adsorption kinetics and higher pesticide adsorption capacities. Density functional theory calculations showed that pesticide adsorption in the Cu-MOFs was mainly electrostatic. A dispersive solid phase extraction method was developed based on a defective Cu-MOF-6 for rapidly extracting pesticides from food samples. The method allowed pesticide detection over a wide linear concentration range, low limits of detection (0.0067-0.0164 µg L-1) and good recoveries in pesticide-spiked samples (81.03-109.55%).

Identification of hub cuproptosis related genes and immune cell infiltration characteristics in periodontitis.

Introduction: Periodontitis is an inflammatory disease and its molecular mechanisms is not clear. A recently discovered cell death pathway called cuproptosis, may related to the disease. Methods: The datasets GSE10334 of human periodontitis and control were retrieved from the Gene Expression Omnibus database (GEO) for analysis.Following the use of two machine learning algorithms, least absolute shrinkage and selection operator (LASSO) and support vector machine-recursive feature removal (SVM-RFE) were used to find CRG-based signature. Then the Receiver operating characteristic (ROC) curves was used to evaluate the gene signature's discriminatory ability. The CIBERSORT deconvolution algorithm was used to study the link between hub genes and distinct types of immune cells. Next, the association of the CRGs with immune cells in periodontitis and relevant clusters of cuproptosis were found. The link between various clusters was ascertained by the GSVA and CIBERSORT deconvolution algorithm. Finally, An external dataset (GSE16134) was used to confirm the diagnosis capacity of the identified biomarkers. In addition, clinical samples were examined using qRT-PCR and immunohistochemistry to verifiy the expression of genes related to cuprotosis in periodontitis and the signature may better predict the periodontitis. Results: 15 periodontitis-related DE-CRGs were found,then 11-CRG-based signature was found by using of LASSO and SVM-RFE. ROC curves also supported the value of signature. CIBERSORT results of immune cell signature in periodontitis showed that signature genes is a crucial component of the immune response.The relevant clusters of cuproptosis found that the NFE2L2, SLC31A1, FDX1,LIAS, DLD, DLAT, and DBT showed a highest expression levels in Cluster2 ,while the NLRP3, MTF1, and DLST displayed the lowest level in Cluster 2 but the highest level in Cluster1. The GSVA results also showed that the 11 cuproptosis diagnostic gene may regulate the periodontitis by affecting immune cells. The external dataset (GSE16134) confirm the diagnosis capacity of the identified biomarkers, and clinical samples examined by qRT-PCR and immunohistochemistry also verified that these cuprotosis related signiture genes in periodontitis may better predict the periodontitis. Conclusion: These findings have important implications for the cuproptosis and periodontitis, and highlight further research is needed to better understand the mechanisms underlying this relationship between the cuproptosis and periodontitis.