Mutual exclusivity and co-occurrence patterns of immune checkpoints indicate NKG2A relates to anti-PD-1 resistance in gastric cancer.

BACKGROUND: An increasing number of clinical studies have begun to explore combination strategies with immune checkpoint inhibitors, aiming to present new opportunities for overcoming anti-PD-1 treatment resistance in gastric cancer. Unfortunately, the exploration of certain immune checkpoint inhibitor combination strategies has yielded suboptimal results. Therefore, it is necessary to comprehensively analyze the expression patterns of immune checkpoints and identify optimal combination regimens of anti-PD-1 inhibitors with other immune checkpoint inhibitors. METHODS: Leveraging single-cell RNA sequencing (scRNA-seq) and multivariate linear regression interaction models, we dissected the immune checkpoint expression characteristics of CD8+ T cells in gastric cancer and the immune checkpoint expression pattern (ICEP) mediating anti-PD-1 treatment resistance. Furthermore, we employed transcription factor analysis and CellOracle to explore the transcriptional regulatory mechanisms governing CD8+ T cell differentiation fates. Finally, we utilized Nichenet and spatial transcriptomic analysis to investigate the spatial expression patterns of immune checkpoints. RESULTS: Interaction analysis indicated that, among the known immune checkpoints, co-expression of NKG2A and PD-1 might exert a more profound inhibitory effect on the proliferative capacity of CD8+ T cells. The co-expression analysis revealed differential co-expression pattern of PD-1 and NKG2A, defined as ICEP1 (CD8+ T cells co-expressing PD-1, CTLA-4, TIGIT, LAG-3 or CD38) and ICEP2 (CD8+ T cells solely expressing NKG2A or co-expressing with other immune checkpoints), reflecting the co-occurrence pattern of PD-1 and the mutual exclusivity of NKG2A. Further, these two ICEP CD8+ T cell subsets represented distinct CD8+ T cell differentiation fates governed by MSC and RUNX3. Notably, ICEP2 CD8+ T cells were associated with anti-PD-1 therapy resistance in gastric cancer. This phenomenon may be attributed to the recruitment of LGMN+ macrophages mediated by the CXCL16-CXCR6 signaling pathway. CONCLUSION: This study unveiled two distinct ICEPs and the mutually exclusivity and co-occurrence characteristics of CD8+ T cells in gastric cancer. The ICEP2 CD8+ T cell subset, highly expressed in gastric cancer patients resistant to anti-PD-1 therapy, may be recruited by LGMN+ macrophages through CXCL16-CXCR6 axis. These findings provide evidence for NKG2A as a novel immunotherapeutic target in gastric cancer and offer new insights into combination strategies for immune checkpoint inhibitors in gastric cancer.

Evodiamine suppresses endometriosis development induced by early EBV exposure through inhibition of ERß.

Introduction: Endometriosis (EMS) is characterized as a prevalent gynecological inflammatory disorder marked by the existence of endometrial tissues situated beyond the uterus. This condition leads to persistent pelvic pain and may contribute to infertility. In this investigation, we explored the potential mechanism underlying the development of endometriosis (EMS) triggered by transient exposure to either latent membrane protein 1 (LMP1) or Epstein-Barr virus (EBV) in a mouse model. Additionally, we examined the potential inhibitory effect of evodiamine (EDM) on EMS. Methods: Immortalized human endometrial stromal cells (HESC) or epithelial cells (HEEC) were transiently exposed to either EBV or LMP1. The presence of evodiamine (EDM) was assessed for its impact on estrogen receptor ß (ERß) expression, as well as on cell metabolism parameters such as redox balance, mitochondrial function, inflammation, and proliferation. Additionally, a mixture of LMP1-treated HESC and HEEC was administered intraperitoneally to generate an EMS mouse model. Different dosages of EDM were employed for treatment to evaluate its potential suppressive effect on EMS development. Results: Transient exposure to either EBV or LMP1 triggers persistent ERß expression through epigenetic modifications, subsequently modulating related cell metabolism for EMS development. Furthermore, 4.0 µM of EDM can efficiently reverse this effect in in vitro cell culture studies. Additionally, 20 mg/kg body weight of EDM treatment can partly suppress EMS development in the in vivo EMS mouse model. Conclusion: Transient EBV/LMP1 exposure triggers permanent ERß expression, favoring later EMS development, EDM inhibits EMS development through ERß suppression. This presents a novel mechanism for the development of endometriosis (EMS) in adulthood stemming from early Epstein-Barr virus (EBV) exposure during childhood. Moreover, evodiamine (EDM) stands out as a prospective candidate for treating EMS.

Pectin from steam explosion-treated citrus peel exhibits good emulsion properties and bioavailability-promoting effect in vitro of nobiletin.

Steam explosion (SE) is a potential method to modify pectin structure, which might be connected to its emulsifying characteristics and the bioavailability of encapsulated polymethoxyflavone like nobiletin. However, the relationship between SE-modified pectin and the bioavailability of encapsulated nobiletin is still unclear. In this study, nobiletin-loaded emulsion was fabricated using citrus pectin modified with SE (0.15-0.9 MPa, 3 min) as emulsifier for in vitro digestion study, and the transport and absorption of nobiletin in Caco-2 cells to investigate the bioavailability-promoting effect. The results showed that SE treatment lowered the droplet size of emulsion from 21.38 ± 2.30 µm to 2.14 ± 0.12 µm, enhanced the nobiletin encapsulation efficiency from 23.73 ± 0.78% to 86.27 ± 3.81%, improved the nobiletin bioaccessibility in vitro from 2.48 ± 0.10% to 25.42 ± 0.10% and increased the intracellular accumulation of nobiletin by over 10 times, even higher than that of Tween 80. In conclusion, pectin from SE-treated citrus peel exhibited good emulsion properties and bioavailability-promoting effect in vitro of nobiletin.

Ixekizumab Improved Refractory Erythrodermic Psoriasis with Comorbid Diffuse Alopecia: A Case Report with 52-Week Follow-Up.

Erythrodermic psoriasis (EP) is a severe and rare variant of psoriasis, accounting for less than 3% of cases. It is characterized by widespread scaling and erythema that affects more than 90% of the body surface area. Alopecia can manifest as a symptom associated with the disease, further exacerbating the impact on the patient's quality of life. We present the case of a patient with severe EP and diffuse alopecia who did not respond to conventional therapies. The patient was subsequently treated with ixekizumab as per labeled usage, resulting in complete resolution of both psoriatic skin lesions (Psoriasis area and severity index/PASI 100) and alopecia (The Severity of Alopecia Tool/SALT 0).

Designing antiperovskite derivatives via atomic-position splitting for photovoltaic applications.

Due to the success of halide perovskites in the photovoltaic field, halide perovskite-derived semiconductors have also been widely studied for optoelectronic applications. However, the photovoltaic performance of these perovskite derivatives still lags significantly behind their perovskite counterparts, mainly due to deficiencies at the B-site or X-site of the derivatives, which disrupt the connectivity of the key [BX6] octahedra units. Herein, we developed a class of antiperovskite-derived materials with the formula , achieved by splitting the A anion, originally at the corner site of the cubic antiperovskite structure, into three edge-centered sites. This structural transformation maintains the three-dimensional octahedral framework. The thermodynamic stability, dynamical stability, and band gaps of 80 compounds were calculated using first-principles calculations. Based on criteria including stability and electronic properties, we identified 9 promising antiperovskite derivatives for further evaluation of their photovoltaic performance. Notably, the calculated theoretical maximum efficiencies of Ba3BiI3, Ba3SbI3, and Ba3BiBr3 all exceed 24.5%, which is comparable to that of CH3NH3PbI3 solar cells. Interpretable machine learning analysis was further carried out to identify critical physical descriptors influencing thermodynamic stability and band gap. Our work provides a novel approach for designing high performance perovskite-type structure-inspired semiconductors with potential for optoelectronic applications.

Design, Synthesis, in Vitro and in Vivo Biological Evaluation of Pterostilbene Derivatives for the Treatment of Inflammatory Bowel Disease.

The aberrant activation of NLRP3 inflammasomes is intricately linked to various inflammatory diseases. In this study, we present the discovery and optimization of a series of NLRP3 inflammasome inhibitors based on the pterostilbene skeleton. All compounds underwent screening to evaluate their inhibitory effects on LPS/Nigericin-induced IL-1ß secretion and anti-cellular pyroptosis. Most compounds exhibit good biological activity and cellular safety, with compound D20 showing the most prominent activity. Preliminary mechanism studies suggest that compound D20 may affect the assembly of NLRP3 inflammasomes by targeting the NLRP3 protein, thereby inhibiting the activation of NLRP3 inflammasomes. The in vivo anti-inflammatory activity demonstrated significant therapeutic effect of compound D20 on DSS-induced acute colitis model in mice. This work has important reference significance for the development of drugs targeting NLRP3 inflammasomes.

Surface inducing high-temperature superconductivity in layered metal carborides Li2BC3 and LiBC by metallizing σ electrons.

Metallizing σ electrons provides a promising route to design high-temperature superconducting materials, such as MgB2 and high-pressure hydrides. Here, we focus on two MgB2-like layered carborides Li2BC3 and LiBC; their bulk does not have superconductivity because the B-C σ states are far away from the Fermi level (EF), however, based on first-principles calculations, we found that when their bulk systems are cleaved into surfaces with B-C termination, high Tc of ∼80 K could be observed in the exposed B-C layer on the surfaces. Detailed analysis reveals that surface symmetry reduction, due to lattice periodic breaking, not only introduces hole self-doping into surface B-C layers and shifts the σ-bonding states towards the EF - associated with emergent large electronic occupation, but also makes in-plane stretching modes on the surface layer experience significant softness. The enhanced σ states and softened phonon modes work to produce strong coupling, thus yielding high-Tc surface superconductivity, which distinctly differs from the superconducting features of the MgB2 film, which generates phonon stiffness accompanied by suppressed superconductivity. Our findings undoubtedly provide a novel platform to realize high-Tc surface superconductivity, and also clearly elucidate the microscopic mechanism of surface-enhanced superconductivity in favor of creating more high-Tc surface superconductors among MgB2-like layered materials.

Cystatin C and sarcopenia index are associated with cardiovascular and all-cause death among adults in the United States.

OBJECTIVES: This study aimed to investigate the association of cystatin C, serum creatinine and sarcopenia index with cardiovascular and all-cause death in general population. METHODS: Data of participants from the National Health and Nutrition Examination Surveys (NHANES) from 1999 to 2004 were used and all participants were followed up regularly until December 31, 2019. Multivariable Cox analysis was used to investigate the association of cystatin C, serum creatinine and sarcopenia index with cardiovascular and all-cause death. Restricted cubic spline was conducted to evaluate the nonlinear association. RESULTS: A total of 9894 participants with a mean age of 45.64 years were enrolled and followed up for a mean duration of 15.62 ± 4.68 years. Approximately 50.3% were male and there were a total of 2681 all-cause deaths and 691 cardiovascular deaths recorded during the follow-up period. In final adjusted model, compared with the first quartile of cystatin C (< 0.659 mg/L), the risk of cardiovascular and all-cause death increased 2.36-fold and 1.71-fold for participants in the fourth quartile (≥ 0.877 mg/L) (HR: 3.36, 95% CI: 2.06-5.46, P < 0.001; HR: 2.71, 95% CI: 2.17-3.38, P < 0.001; respectively). Furthermore, a higher sarcopenia index (< 88.41 vs. ≥125.52) was associated with the reduced risk of cardiovascular death (HR: 0.41, 95% CI: 0.31-0.53, P < 0.001) as well as all-cause death (HR: 0.41, 95% CI: 0.35-0.49, P < 0.001). Additionally, restricted cubic splines showed that there was a nonlinear relationship between sarcopenia index levels and all-cause death while there was a linear relationship between sarcopenia index levels and cardiovascular death. CONCLUSIONS: Higher sarcopenia index was associated with the decreased risk of cardiovascular and all-cause death in general population in the United States. Elevated cystatin C was positively associated with cardiovascular and all-cause death.

Groundwater flow regime shapes nitrogen functional traits by affecting microbial community assembly processes in the subsurface.

The complex nitrogen (N) cycle in groundwater systems is affected by both biological and environmental factors. The interactions between hydrogeological conditions and the microbial community assembly processes that impact N-cycling processes remain poorly understood. We explored the assembly patterns of N-cycling microbial communities along the groundwater flow path. The environmental heterogeneity in different hydrological phases increased along the flow path (mean Ed: 0.16-0.49), accompanied by different microbial community assembly patterns. The assembly patterns that engaged in dissimilatory nitrate reduction to ammonium (DNRA) and denitrification changed across the water-sediment phases. Nitrifying microorganisms in the discharge area were mainly influenced by heterogeneous selection (41-69 %), and were closely correlated with dissolved oxygen (DO) concentrations. Homogeneity along flow-through increased stochastic assemblies, such as downstream drift of anammox bacterial (AnAOB) communities. Thus, the N removal pathway changed from "nitrification-denitrification" in the recharge area to "partial nitrification-anammox" in the discharge area. The increasing environmental heterogeneity brought more deterministic assembly patterns of N-cycling communities, linked to higher community turnover along the groundwater flow path. This study indicated that groundwater flow regime determined microbial community assembly patterns, providing valuable insight into the response of N transitions to environmental variations in groundwater systems.

PE6c greatly enhances prime editing in transgenic rice plants.

Prime editing is a versatile CRISPR/Cas-based precise genome-editing technique for crop breeding. Four new types of prime editors (PEs) named PE6a-d were recently generated using evolved and engineered reverse transcriptase (RT) variants from three different sources. In this study, we tested the editing efficiencies of four PE6 variants and two additional PE6 constructs with double-RT modules in transgenic rice (Oryza sativa) plants. PE6c, with an evolved and engineered RT variant from the yeast Tf1 retrotransposon, yielded the highest prime-editing efficiency. The average fold change in the editing efficiency of PE6c compared with PEmax exceeded 3.5 across 18 agronomically important target sites from 15 genes. We also demonstrated the feasibility of using two RT modules to improve prime-editing efficiency. Our results suggest that PE6c or its derivatives would be an excellent choice for prime editing in monocot plants. In addition, our findings have laid a foundation for prime-editing-based breeding of rice varieties with enhanced agronomically important traits.

The Anti-Biofilm Properties of Phloretin and Its Analogs against Porphyromonas gingivalis and Its Complex Flora.

Porphyromonas gingivalis is crucial for the pathogenesis of periodontitis. This research investigated the effects of the fruit-derived flavonoid phloretin and its analogs on the growth of pure P. gingivalis and the flora of P. gingivalis mixed with the symbiotic oral pathogens Fusobacterium nucleatum and Streptococcus mitis. The results showed that the tested flavonoids had little effect on the biofilm amount of pure P. gingivalis, but significantly reduced the biofilm amount of mixed flora to 83.6~89.1%. Biofilm viability decreased to 86.7~92.8% in both the pure- and mixed-bacterial groups after naringenin and phloretin treatments. SEM showed that phloretin and phlorizin displayed a similar and remarkable destructive effect on P. gingivalis and the mixed biofilms. Transcriptome analysis confirmed that biofilm formation was inhibited by these flavonoids, and phloretin significantly regulated the transcription of quorum sensing. Phlorizin and phloretin reduced AI-2 activity to 45.9% and 55.4%, respectively, independent of the regulation of related gene transcription. This research marks the first finding that these flavonoids possess anti-biofilm properties against P. gingivalis and its intricate bacterial community, and the observed performance variations, driven by structural differences, underscore the existence of intriguing structure-activity relationships.

Lysosomal-Associated Protein Transmembrane 5, Tubular Senescence, and Progression of CKD.

BACKGROUND: Tubular senescence is a major determinant of chronic kidney disease (CKD) and identification of potential therapeutic targets involved in senescent tubular epithelial cells has clinical importance. Lysosomal-associated protein transmembrane 5 (LAPTM5) is a key molecule related to T and B cell receptor expression and inflammation. However, the expression pattern of LAPTM5 in the kidney and the contribution of LAPTM5 to the development of CKD keep unknown. METHODS: LAPTM5-/- mice and tubule specific-LAPTM5 knockout mice were used to examine the role of LAPTM5 in tubular senescence by establishing different experimental mouse CKD models. RESULTS: LAPTM5 expression was significantly induced in the kidney, especially in proximal tubules and distal convoluted tubules, from mice with aristolochic acid nephropathy, bilateral ischemia/reperfusion injury (IRI)-induced CKD or unilateral ureter obstruction (UUO). Tubule-specific deletion of LAPTM5 inhibited senescence of tubular epithelial cells and alleviated tubulointerstitial fibrosis in aged mice. Moreover, LAPTM5 deficiency ameliorated kidney injury and tubular senescence in mice with CKD. Mechanistically, LAPTM5 inhibited ubiquitination of NICD1 by mediating WWP2 lysosomal degradation, then leading to cellular senescence in tubular epithelial cells. Notably, we also observed a higher expression of LAPTM5 in tubules from individuals with CKD and the level of LAPTM5 was correlated with kidney fibrosis and tubular senescence in people with CKD. CONCLUSIONS: LAPTM5 contributed to tubular senescence by regulating WWP2/NICD1 signaling pathway and exacerbated kidney injury during the progression of CKD.

Prognostic and clinicopathological significance of FOXD1 in various cancers: a meta and bioinformation analysis.

Aim: To examine both predictive and clinicopathological importance underlying FOXD1 in malignant tumors, our study adopts meta-analysis. Methods: We searched from PubMed, Embase, WOS, Wanfang and CNKI. Stata SE15.1 was used to calculate the risk ratio (HR) as well as relative risk (RR) with 95% of overall CIs to assess FOXD1 and overall survival rate (OS), disease-free survival rate as well as clinicopathological parameters. Results: 3808 individuals throughout 17 trials showed high FOXD1 expression was linked to disadvantaged OS (p < 0.001) and disease-free survival (p < 0.001) and higher TNM stage (p < 0.001). Conclusion: Elevated FOXD1 had worse predictions and clinicopathological parameters in most cancers. The GEPIA database findings also support our results.

FOXD1 is a gene linked to a variety of cancers. In our article, we analyzed the results of several clinical trials in patients with different cancers. We found that when this gene is expressed in large amounts, it is often indicative of poor survival rates. From this study we can use FOXD1 to predict the course of the disease and at the same time study its upper and lower pathways to find therapeutic drugs to treat the cancer.

Minimal Common Oncology Data Elements Genomics Pilot Project: Enhancing Oncology Research Through Electronic Health Record Interoperability at Vanderbilt University Medical Center.

PURPOSE: The expanding presence of the electronic health record (EHR) underscores the necessity for improved interoperability. To test the interoperability within the field of oncology research, our team at Vanderbilt University Medical Center (VUMC) enabled our Epic-based EHR to be compatible with the Minimal Common Oncology Data Elements (mCODE), which is a Fast Healthcare Interoperability Resources (FHIR)-based consensus data standard created to facilitate the transmission of EHRs for patients with cancer. METHODS: Our approach used an extract, transform, load tool for converting EHR data from the VUMC Epic Clarity database into mCODE-compatible profiles. We established a sandbox environment on Microsoft Azure for data migration, deployed a FHIR server to handle application programming interface (API) requests, and mapped VUMC data to align with mCODE structures. In addition, we constructed a web application to demonstrate the practical use of mCODE profiles in health care. RESULTS: We developed an end-to-end pipeline that converted EHR data into mCODE-compliant profiles, as well as a web application that visualizes genomic data and provides cancer risk assessments. Despite the complexities of aligning traditional EHR databases with mCODE standards and the limitations of FHIR APIs in supporting advanced statistical methodologies, this project successfully demonstrates the practical integration of mCODE standards into existing health care infrastructures. CONCLUSION: This study provides a proof of concept for the interoperability of mCODE within a major health care institution's EHR system, highlighting both the potential and the current limitations of FHIR APIs in supporting complex data analysis for oncology research.

Molecular structural modification of myofibrillar protein from oyster (Crassostrea gigas) with oligosaccharides for improving its gel properties.

Glycation is a promising approach to enhance protein gel characteristics in the food industry. The impact of oyster myofibrillar protein (MP) being glycosylated with six oligosaccharides (dextran [Dex]-1 kDa, 5 kDa, 6 kDa, and 10 kDa, xylan [Xyla], and xyloglucan [Xyg]) on structural properties, aggregation behavior and gel properties was investigated in this study. The findings demonstrated that oligosaccharides significantly increased the glycation degree of MP by forming a stable tertiary conformation, increasing the contents of the disulfide bond and hydrogen bonds. Additionally, particle sizes decreased and solubility increased after glycation, improving the gel's strength, water-holding capacity, thermal stability, elastic modulus, and ordered network layout. It was determined that MP-Dex 5 had the best gel properties. The gel strength and water holding capacity of MP-Dex 5 increased by 70.59% and 32.27%, respectively. Molecular dynamics simulations results showed van der Waals energy and electrostatic interactions favor myosin binding to Dex or Xyla units. This study will provide insights into the relationship between molecular structure, aggregation behavior and gel property of oyster MP-oligosaccharide couples, and expand the application of oyster MP in food gels.

Could school climate moderate the relationship between school bullying patterns and academic burnout in Chinese adolescents: A large cross-sectional study.

OBJECTIVES: Students who are bullied not only affect academic performance, but also produce a range of psychological problems. The purpose of the present study was to investigate the association between school bullying and academic burnout among Chinese students, assuming school climate to play a moderating role in the aforementioned relationship. This study provides corresponding intervention strategies and reference data for the prevention and treatment of bullying in schools. METHODS: The data was obtained by a cross-sectional survey of 20,730 Chinese students from 23rd May to 23rd June 2022. Multiple linear regressions and Latent Profile Analysis (LPA) were used to examine the hypotheses. RESULTS: This study revealed that all dimensions of school bullying and school bullying level (ß = -0.09; 95 % CI, -4.946, -3.833) were associated with academic burnout. Verbal bullying (ß = 0.15; 95 % CI, 1.689, 1.972) had the strongest association with academic burnout, followed by social (ß = 0.14; 95 % CI, 1.496, 1.779) and physical bullying (ß = 0.13; 95 % CI, 1.451, 1.734), while cyber bullying (ß = 0.08; 95 % CI, 0.847, 1.127) had the weakest association with academic burnout. In addition, school climate can moderate the association between school bullying and academic burnout. Students who experienced school bullying and in bad school climate showed elevated levels of academic. LIMITATIONS: This study used cross-sectional data, preventing us from drawing conclusions about causation. CONCLUSIONS: The findings suggested that creating a harmonious school climate and reducing school bullying may effectively alleviate academic burnout caused by school climate and school bullying.

High-mobility InSnZnO Thin Film Transistors via Introducing Water Vapor Sputtering Gas.

There is always a doubt that introducing water during oxide growing has a positive or negative effect on the properties of oxide films and devices. Herein, a comparison experiment on the condition of keeping the same oxygen atom flux in the sputtering chamber is designed to examine the influences of H2O on In-Sn-Zn-O (ITZO) films and their transistors. In comparison to no-water films, numerous unstable hydrogen-related defects are induced on with-water films at the as-deposited state. Paradoxically, this induction triggers an ordered enhancement in the microstructure of the films during conventional annealing, characterized by a reduction in H-related and vacancy (Vo) defects as well as an increase in film packing density and the M-O network ordering. Ultimately, the no-water thin-film transistors (TFTs) exhibit nonswitching behavior, whereas 5 sccm-water TFT demonstrates excellent electrical performance with a remarkable saturation field-effect mobility (µFE) of 122.10 ± 5.00 cm2·V-1·s-1, a low threshold (Vth) of -2.30 ± 0.40 V, a steep sub-threshold swing (SS) of 0.18 V·dec-1, a high output current (Ion) of 1420 µA, and a small threshold voltage shift ΔVth of -0.77 V in the negative bias stability test (3600 s).

Single-cell transcriptomics across 2,534 microbial species reveals functional heterogeneity in the rumen microbiome.

Deciphering the activity of individual microbes within complex communities and environments remains a challenge. Here we describe the development of microbiome single-cell transcriptomics using droplet-based single-cell RNA sequencing and pangenome-based computational analysis to characterize the functional heterogeneity of the rumen microbiome. We generated a microbial genome database (the Bovine Gastro Microbial Genome Map) as a functional reference map for the construction of a single-cell transcriptomic atlas of the rumen microbiome. The atlas includes 174,531 microbial cells and 2,534 species, of which 172 are core active species grouped into 12 functional clusters. We detected single-cell-level functional roles, including a key role for Basfia succiniciproducens in the carbohydrate metabolic niche of the rumen microbiome. Furthermore, we explored functional heterogeneity and reveal metabolic niche trajectories driven by biofilm formation pathway genes within B. succiniciproducens. Our results provide a resource for studying the rumen microbiome and illustrate the diverse functions of individual microbial cells that drive their ecological niche stability or adaptation within the ecosystem.

Differential expression of sNPF in male and female eyestalk leading to sex dimorphism of AMP expression in Procambarus clarkii intestine.

Antimicrobial peptide (AMP) is an important component of crustaceans' innate immune system. In this study, a short neuropeptide F (sNPF) gene (Pc-sNPF) and a Forkhead box O (FOXO) gene (PcFOXO) from Procambarus clarkii were identified. Analysis findings showed that the expression level of AMP genes differed between male and female P. clarkii. Furthermore, Pc-sNPF and PcFOXO were related to the sex dimorphism of AMP. Knockdown of Pc-sNPF in the eyestalk significantly upregulated the expression of PcFOXO and two anti-lipopolysaccharide factors (PcALF4 and PcALFL) in the intestine of P. clarkii. The expression of PcFOXO in the intestine of female P. clarkii was higher than in that of males. Results from RNA interference revealed that PcFOXO positively regulated the expression of PcALF4 and PcALFL in the intestine of male and female P. clarkii. In summary, our study showed that differences in Pc-sNPF expression in eyestalk of male and female P. clarkii leading to sex dimorphism of AMP expression in the intestine are mediated by the sNPF-FOXO-AMP signal pathway called the eyestalk-intestine axis.

BNT12, a novel hybrid peptide of opioid and neurotensin pharmacophores, produces potent central antinociception with limited side effects.

The development of multitarget opioid drugs has emerged as an attractive approach for innovative pain management with reduced side effects. In the present study, a novel hybrid peptide BNT12 containing the opioid and neurotensin (NT)-like fragments was synthesized and pharmacologically characterized. In acute radiant heat paw withdrawal test, intracerebroventricular (i.c.v.) administration of BNT12 produced potent antinociception in mice. The central antinociceptive activity of BNT12 was mainly mediated by µ-, δ-opioid receptor, neurotensin receptor type 1 (NTSR1) and 2 (NTSR2), supporting a multifunctional agonism of BNT12 in the functional assays. BNT12 also exhibited significant antinociceptive effects in spared nerve injury (SNI)-neuropathic pain, complete Freund's adjuvant (CFA)-induced inflammatory pain, acetic acid-induced visceral and formalin-induced pain after i.c.v. administration. Furthermore, BNT12 exhibited substantial reduction of acute antinociceptive tolerance, shifted the dose-response curve to the right by only 1.3-fold. It is noteworthy that BNT12 showed insignificant chronic antinociceptive tolerance at the supraspinal level. In addition, BNT12 exhibited reduced or no opioid-like side effects on conditioned place preference (CPP) response, naloxone-precipitated withdrawal response, acute hyperlocomotion, motor coordination, gastrointestinal transit, and cardiovascular responses. The present investigation demonstrated that the novel hybrid peptide BNT12 might serve as a promising analgesic candidate with limited opioid-like side effects.