Penisimplinoids A-K, highly oxygenated andrastin-type meroterpenoids with diverse activities from the marine-derived fungus Penicillium simplicissimum.

Penisimplinoid A (1), the first andrastin-type meroterpenoid with an unprecedented 6/6/3/6/5/5 polycyclic systems, together with ten highly oxygenated andrastin-type meroterpenoids (2-11) and one known analogue (12), were co-isolated from the marine-derived fungus Penicillium simplicissimum. Their absolute configurations were determined by single-crystal X-ray diffraction analysis (Cu Kα), DP4+ probability analyses, and ECD quantum chemistry calculations. Biological evaluation revealed that 7 and 12 showed anti-inflammatory activities in the zebrafish assay, 6 exhibited cytotoxic activity against NCI-H446 tumor cells with an IC50 value of 6.49 µM, 7 and 11 exhibited significant promoting angio-genesis activities.

A diverse Fusarium community is responsible for contamination of rice with a variety of Fusarium toxins.

Rice plays an important role in the daily diet in China and therefore its quality and safety have been of great concern. However, few systematic studies have investigated Fusarium community and toxins in rice grains. Here, we collected 1381 rice samples from Jiangsu Province in eastern China and found a higher frequency of zearalenone (ZEN), deoxynivalenol (DON), fumonisins (FBs), and beauvericin (BEA). The positive samples were individually contaminated with a minimum of one and a maximum of ten toxins. Fusarium was isolated and identified as the major fungus, which exhibited temporal and geographical distribution. The most prevalent species complexes within this genus were Fusarium incarnatum-equiseti species complex (FIESC), Fusarium fujikuroi species complex (FFSC), and Fusarium sambucinum species complex (FSAMSC). Nevertheless, the amplicon sequence analysis revealed a low relative abundance of Fusarium in the rice panicles, and the fungal community exhibited an irregular change along with the symptom's emergence. In vitro toxigenic profiles of Fusarium strains showed significant complexity and specificity depending on the type and content. FIESC strains were non-pathogenic to wheat heads and weakly pathogenic to maize ears, respectively, accumulating lower amounts of toxins than F. asiaticum and F. fujikuroi. There was no significant variation in the ability to cause panicle blight in rice among the various species tested. Our study provides detailed information about the contamination of Fusarium toxins and community in rice after harvest. This information is valuable for understanding the relationship between Fusarium and rice and for developing effective control strategies.

Altered metabolism in cancer: insights into energy pathways and therapeutic targets.

Cancer cells undergo significant metabolic reprogramming to support their rapid growth and survival. This study examines important metabolic pathways like glycolysis, oxidative phosphorylation, glutaminolysis, and lipid metabolism, focusing on how they are regulated and their contributions to the development of tumors. The interplay between oncogenes, tumor suppressors, epigenetic modifications, and the tumor microenvironment in modulating these pathways is examined. Furthermore, we discuss the therapeutic potential of targeting cancer metabolism, presenting inhibitors of glycolysis, glutaminolysis, the TCA cycle, fatty acid oxidation, LDH, and glucose transport, alongside emerging strategies targeting oxidative phosphorylation and lipid synthesis. Despite the promise, challenges such as metabolic plasticity and the need for combination therapies and robust biomarkers persist, underscoring the necessity for continued research in this dynamic field.

A self-healing, long-lasting adhesive, lignin-based polyvinyl alcohol organo-hydrogel for strain-sensing applications.

Hydrogel-based flexible sensors have garnered considerable interest in the fields of soft electronics, robotics, and human-machine interfaces. For better practical applications, integrating multiple properties-such as self-adhesive, anti-freeze, anti-volatile, self-healing, and antibacterial-into a single gel for flexible sensors remains a challenge. In this paper, a multifunctional lignin-based polyvinyl alcohol gel, containing dynamic covalent bonds, hydrogen bonds, and coordination bonds, is constructed by a simple one-pot method, in which ethylene glycol/water chosen as a binary solvent and KI as a conductive medium. The resulting organogel exhibits self-healing, long-lasting adhesion, UV shielding, antibacterial properties, excellent frost resistance (-20 °C), and volatile resistance properties. In addition, the organogel-based sensor demonstrates satisfactory sensitivity in detecting joint movements and facial expressions. This study provides a new strategy for developing a versatile flexible sensor through the introduction of renewable and bio-based lignin, promising applications in the fields of wearable electronics.

Chromosome-scale genome assembly of Docynia delavayi provides new insights into the α-farnesene biosynthesis.

Docynia delavayi is an economically significant fruit species with a high market potential due to the special aroma of its fruit. Here, a 653.34 Mb high-quality genome of D. delavayi was first reported, of which 93.8 % of the sequences (612.98 Mb) could be anchored to 17 chromosomes, containing 48,325 protein-coding genes. Ks analysis proved that two whole genome duplication (WGD) events occurred in D. delavayi, resulting in the expansion of genes associated with terpene biosynthesis, which promoted its fruit-specific aroma production. Combined multi-omics analysis, α-farnesene was detected as the most abundant aroma substance emitted by D. delavayi fruit during storage, meanwhile one α-farnesene synthase gene (AFS) and 15 transcription factors (TFs) were identified as the candidate genes potentially involved in α-farnesene biosynthesis. Further studies for the regulation network of α-farnesene biosynthesis revealed that DdebHLH, DdeERF1 and DdeMYB could activate the transcription of DdeAFS. To our knowledge, it is the first report that MYB TF plays a regulatory role in α-farnesene biosynthesis, which will greatly facilitate future breeding programs for D. delavayi.

Cytochrome P450 F3 promotes colorectal cancer via inhibiting NRF2-mediated ferroptosis.

Cytochrome P450 F3 (CYP4F3) is recognized as a disease-associated immune response initiator that is involved in the synthesis of cholesterol, steroids, and lipids. This study identified the upregulation of CYP4F3 expression in colorectal cancer (CRC) and its association with poor patient prognosis through a comparative analysis between CRC tumor tissues with normal tissues from public databases. The overexpression of CYP4F3 in CT26.wt and SW620, promoted cell proliferation and migration, a reduction of cellular oxidative stress, an up-regulation of the oxidative stress-related pathway NRF2, and an inhibition of cellular ferroptosis. Additionally, inhibition of NRF2 activity stimulated cellular ferroptosis when CYP4F3 was overexpressed. Ferroptosis, characterized by iron-dependent lipid peroxidation, is a non-apoptotic way of cell death with a critical role in cancer development. When given a ferroptosis agonist to CYP4F3-overexpression CRC cells, NRF2 was activated, and cell proliferation and migration were reduced. Furthermore, the mice subcutaneously injected with CYP4F3-overexpression CT26.wt cells formed significantly larger tumors compared to the CYP4F3-vector CT26.wt cell group. This study systematically identified an important role of CYP4F3 in CRC development as a regulator of CRC cells to escape ferroptosis via NRF2, highlighting the significance of CYP4F3 as a potential therapeutic target for CRC.

Hallmarks of cancer resistance.

This review explores the hallmarks of cancer resistance, including drug efflux mediated by ATP-binding cassette (ABC) transporters, metabolic reprogramming characterized by the Warburg effect, and the dynamic interplay between cancer cells and mitochondria. The role of cancer stem cells (CSCs) in treatment resistance and the regulatory influence of non-coding RNAs, such as long non-coding RNAs (lncRNAs), microRNAs (miRNAs), and circular RNAs (circRNAs), are studied. The chapter emphasizes future directions, encompassing advancements in immunotherapy, strategies to counter adaptive resistance, integration of artificial intelligence for predictive modeling, and the identification of biomarkers for personalized treatment. The comprehensive exploration of these hallmarks provides a foundation for innovative therapeutic approaches, aiming to navigate the complex landscape of cancer resistance and enhance patient outcomes.

Closed-loop recycling of tough epoxy supramolecular thermosets constructed with hyperbranched topological structure.

The regulation of topological structure of covalent adaptable networks (CANs) remains a challenge for epoxy CANs. Here, we report a strategy to develop strong and tough epoxy supramolecular thermosets with rapid reprocessability and room-temperature closed-loop recyclability. These thermosets were constructed from vanillin-based hyperbranched epoxy resin (VanEHBP) through the introduction of intermolecular hydrogen bonds and dual dynamic covalent bonds, as well as the formation of intramolecular and intermolecular cavities. The supramolecular structures confer remarkable energy dissipation capability of thermosets, leading to high toughness and strength. Due to the dynamic imine exchange and reversible noncovalent crosslinks, the thermosets can be rapidly and effectively reprocessed at 120 °C within 30 s. Importantly, the thermosets can be efficiently depolymerized at room temperature, and the recovered materials retain the structural integrity and mechanical properties of the original samples. This strategy may be employed to design tough, closed-loop recyclable epoxy thermosets for practical applications.

Cellular Aging and Senescence in Cancer: A Holistic Review of Cellular Fate Determinants.

This comprehensive review navigates the complex relationship between cellular aging, senescence, and cancer, unraveling the determinants of cellular fate. Beginning with an overview of cellular aging's significance in cancer, the review explores processes, changes, and molecular pathways influencing senescence. The review explores senescence as a dual mechanism in cancer, acting as a suppressor and contributor, focusing on its impact on therapy response. This review highlights opportunities for cancer therapies that target cellular senescence. The review further examines the senescence-associated secretory phenotype and strategies to modulate cellular aging to influence tumor behavior. Additionally, the review highlights the mechanisms of senescence escape in aging and cancer cells, emphasizing their impact on cancer prognosis and resistance to therapy. The article addresses current advances, unexplored aspects, and future perspectives in understanding cellular aging and senescence in cancer.

Detection of Multi-Layered Bond Delamination Defects Based on Full Waveform Inversion.

This study aimed to address the challenges encountered in traditional bulk wave delamination detection methods characterized by low detection efficiency. Additionally, the limitations of guided wave delamination detection methods were addressed, particularly those utilizing reflected waves, which are susceptible to edge reflections, thus complicating effective defect extraction. Leveraging the full waveform inversion algorithm, an innovative approach was established for detecting delamination defects in multi-layered structures using ultrasonic guided wave arrays. First, finite element modeling was employed to simulate guided wave data acquisition by a circular array within an aluminum-epoxy bilayer structure with embedded delamination defects. Subsequently, the full waveform inversion algorithm was applied to reconstruct both regular and irregular delamination defects. Analysis results indicated the efficacy of the proposed approach in accurately identifying delamination defects of varying shapes. Furthermore, an experimental platform for guided wave delamination defect detection was established, and experiments were conducted on a steel-cement bilayer structure containing an irregular delamination defect. The experimental results validated the exceptional imaging precision of our proposed technique for identifying delamination defects in multi-layered boards. In summary, the proposed method can accurately determine both the positions and sizes of defects with higher detection efficiency than traditional pulse-echo delamination detection methods.

Assessing health behavior change and comparing remote, hybrid and in-person implementation of a school-based health promotion and coaching program for adolescents from low-income communities.

To assess the impact of a school-based health intervention on adolescents' health knowledge, psychosocial assets and health behaviors, including comparisons of implementation mode: remote, hybrid or in-person. The Stanford Youth Diabetes Coaches Program, an 8-week, school-based health promotion and coaching skills program, was offered to adolescents (ages 14-18 years) from four low-income US communities. Mode of program implementation was remote, hybrid or in-person. Participants completed online pre- and postsurveys. Analysis included paired t-tests, linear regression and qualitative coding. From Fall 2020 to Fall 2021, 262 adolescents enrolled and 179 finished the program and completed pre- and postsurveys. Of the 179, 80% were female, with a mean age of 15.9 years; 22% were Asian; 8% were Black or African American; 25% were White; and 40% were Hispanic. About 115 participants were remote, 25 were hybrid and 39 were in-person. Across all participants, significant improvements (P < 0.01) were reported in health knowledge, psychosocial assets (self-esteem, self-efficacy and problem-solving) and health behaviors (physical activity, nutrition and stress reduction). After adjusting for sex and age, these improvements were roughly equivalent across the three modes of delivery. Participation was associated with significant improvements in adolescent health behaviors. Furthermore, remote mode of instruction was just as effective as in-person and hybrid modes.

Biomimetic Electronic Skin Based on a Stretchable Ionogel Mechanoreceptor Composed of Crumpled Conductive Rubber Electrodes for Synchronous Strain, Pressure, and Temperature Detection.

Electronic skin (e-skin) is showing a huge potential in human-computer interaction, intelligent robots, human health, motion monitoring, etc. However, it is still challenging for e-skin to realize distinguishable detection of stretching strain, vertical pressure, and temperature through a simple noncoupling structure design. Here, a stretchable multimodal biomimetic e-skin was fabricated by integrating layer-by-layer self-assembled crumpled reduced graphene oxide/multiwalled carbon nanotubes film on natural rubber (RGO/MWCNTs@NR) as stretchable conductive electrodes and polyacrylamide/NaCl ionogel as a dielectric layer into an ionotropic capacitive mechanoreceptor. Unlike natural skin receptors, the sandwich-like stretchable ionogel mechanoreceptor possessed a distinct ionotropic capacitive behavior for strain and pressure detection. The results showed that the biomimetic e-skin displayed a negative capacitance change with superior stretchability (0-300%) and a high gauge factor of 0.27 in 180-300% strain, while exhibiting a normal positive piezo-capacitance behavior in vertical pressure range of 0-15 kPa with a maximal sensitivity of 1.759 kPa-1. Based on this feature, the biomimetic e-skin showed an excellent synchronous detection capability of planar strain and vertical pressure in practical wearable applications such as gesture recognition and grasping movement detection without a complicated mathematical or signal decoupling process. In addition, the biomimetic e-skin exhibited a quantifiable linear responsiveness to temperature from 20-90 °C with a temperature coefficient of 0.55%/°C. These intriguing properties gave the biomimetic e-skin the ability to perform a complete function similar to natural skin but beyond its performance for future wearable devices and artificial intelligence devices.

Metagenomic insights into the mechanism for the rapid enrichment and high stability of Candidatus Brocadia facilitated by Fe(â ¢).

The rapid enrichment of anammox bacteria and its fragile resistance to adverse environment are the critical problems facing of anammox processes. As an abundant component in anammox bacteria, iron has been proved to promote the activity and growth of anammox bacteria in the mature anammox systems, but the functional and metabolic profiles in Fe(III) enhanced emerging anammox systems have not been evaluated. Results indicated that the relative abundance of functional genes involved in oxidative phosphorylation, nitrogen metabolism, cofactors synthesis, and extracellular polymers synthesis pathways was significantly promoted in the system added with 5 mg/L Fe(III) (R5). These enhanced pathways were crucial to energy generation, nitrogen removal, cell activity and proliferation, and microbial self-defense, thereby accelerating the enrichment of anammox bacteria Ca. Brocadia and facilitating their resistance to adverse environments. Microbial community analysis showed that the proportion of Ca. Brocadia in R5 also increased to 64.42 %. Hence, R5 could adapt rapidly to the increased nitrogen loading rate and increase the nitrogen removal rate by 108 % compared to the system without Fe(III) addition. However, the addition of 10 and 20 mg/L Fe(III) showed inhibitory effects on the growth and activity of anammox bacteria, which exhibited the lower relative abundance of Ca. Brocadia and unstable or even collapsed nitrogen removal performance. This study not only clarified the concentration range of Fe(III) that promoted and inhibited the enrichment of anammox bacteria, but also deepened our understanding of the functional and metabolic mechanisms underlying enhanced enrichment of anammox bacteria by Fe(III), providing a potential strategy to hasten the start-up of anammox from conventional activated sludge.

Efficacy and Safety of Epidural Chloroprocaine for Breakthrough Pain During Labor Analgesia: A Prospective, Double-Blind, Randomized Trial.

INTRODUCTION: A significant number of women who undergo neuraxial labor analgesia experience breakthrough pain. Prompt mitigation of breakthrough pain is essential to improve maternal and fetal outcomes. We evaluated epidural chloroprocaine compared with ropivacaine in alleviating labor breakthrough pain. METHODS: We performed a double-blind randomized controlled clinical trial between May and July 2023. Eligible parturients received epidural analgesia with ropivacaine and sufentanil. Those with breakthrough pain were randomized to receive either 0.125% epidural ropivacaine (group R) or chloroprocaine at concentrations of 0.5% (group C1), 1.0% (group C2), or 1.5% (group C3), all in a volume of 6 mL. The primary outcome was the treatment success rate, indicated by a decrease of at least 4 points on the numerical rating scale pain score 9 min after analgesic injection. Secondary outcomes and adverse effects were also recorded. RESULTS: Out of 323 patients receiving epidural analgesia, 192 experienced breakthrough pain. After exclusion of three patients because of protocol deviation, there were 47, 48, 47, and 47 patients in group R, C1, C2, and C3, respectively. Group C3 demonstrated a higher treatment success rate (39/47, 83.0%) in managing breakthrough pain than group R (26/47, 55.3%), group C1 (12/48, 25.0%), and group C2 (30/47, 63.8%) (p < 0.001). Group C3 had lower numerical rating scale scores at 6 and 9 min after injection and required fewer patient-controlled epidural boluses than other groups. In addition, group C3 reported greater satisfaction than the other groups (p < 0.001). No significant differences were observed in obstetric or neonatal outcomes across these groups. CONCLUSION: Parturients experiencing breakthrough pain could receive 1.5% epidural chloroprocaine, rather than lower chloroprocaine concentrations and ropivacaine, to achieve more rapid and better pain relief with higher patient satisfaction. TRIAL REGISTRATION: Chinese Clinical Trial Registry, ChiCTR2300071069, http://www.chictr.org.cn/index.aspx .

Predictive, preventive, and personalized medicine in breast cancer: targeting the PI3K pathway.

Breast cancer (BC) is a multifaceted disease characterized by distinct molecular subtypes and varying responses to treatment. In BC, the phosphatidylinositol 3-kinase (PI3K) pathway has emerged as a crucial contributor to the development, advancement, and resistance to treatment. This review article explores the implications of the PI3K pathway in predictive, preventive, and personalized medicine for BC. It emphasizes the identification of predictive biomarkers, such as PIK3CA mutations, and the utility of molecular profiling in guiding treatment decisions. The review also discusses the potential of targeting the PI3K pathway for preventive strategies and the customization of therapy based on tumor stage, molecular subtypes, and genetic alterations. Overcoming resistance to PI3K inhibitors and exploring combination therapies are addressed as important considerations. While this field holds promise in improving patient outcomes, further research and clinical trials are needed to validate these approaches and translate them into clinical practice.

Associations of different isomeric forms of serum lycopene with cardiovascular disease and all-cause mortality.

Background: The effect of serum lycopene on the progression of cardiovascular diseases (CVDs) and their longevity remains a controversial topic. The purpose of this study was to evaluate the associations of different isomeric forms of serum lycopene with CVD and all-cause mortality in the American population. Methods: The National Health and Nutrition Examination Survey (NHANES) is a large population survey to investigate public health in the US. We analyzed data from 2003-2006 linked with mortality data obtained in 2015. Cox proportional hazard ratios (HRs) with 95% confidence intervals (CIs) were estimated to assess the risk of CVD and all-cause mortality caused by serum lycopene. Results: Among 7452 participants (aged 20-85 years, 46.7% male), 298 died from CVDs among the total 1213 deaths during a median follow-up of 10.7 years. Serum lycopene is a protective factor for all-cause and CVD mortality. In multivariable-adjusted models, the hazard ratio (with 95% confidence intervals) associated with Q4 compared to Q1 of serum total-lycopene, trans-lycopene and cis-lycopene was 0.49 (0.38,0.63), 0.49 (0.39,0.63) and 0.55 (0.43,0.70) for all-cause mortality (Ptrend<0.05), and was 0.53 (0.32,0.96), 0.48 (0.32,0.72) and 0.63 (0.41,0.97) for CVD mortality (Ptrend<0.05). The subgroup analyses showed that different isomeric forms of lycopene showed varied associations with CVD and all-cause mortality based on age, drinking status, history of hypertension and diabetes. Conclusions: Serum lycopene concentration was significantly associated with the risk of CVD and all-cause mortality. Cis-lycopene had a U-shaped relationship with mortality, while trans-lycopene had an inverse relationship with it.

High-Precision Corrosion Detection via SH1 Guided Wave Based on Full Waveform Inversion.

Corrosion detection for industrial settings is crucial for safe and efficient operations. Due to its high imaging resolution, the guided-wave full-waveform inversion tomography technique has significant potential for corrosion detection of plate metals. Limited by the long wavelengths of A0 and S0 mode waves, this method exhibits inadequate detection resolution for the earlier shallow and small corrosion defects. Based on the relatively short wavelength characteristics of the SH1 mode wave, we propose a high-precision corrosion detection method via SH1 guided wave using the full waveform inversion algorithms. By conducting finite element simulations of ultrasonic-guided waves on aluminum plates with varying corrosion defects, a comparison was made to assess the detection precision across A0, S0, and SH1 modes. The comparison results showed that, whether for regular or irregular defects, the SH1 mode wave always exhibited higher imaging accuracy than the A0 and S0 mode waves for shallow and small-sized defects. The corresponding experiments were conducted on an aluminum plate with simple or complex defects. The results of the experiments reconfirmed that the full waveform inversion method using SH1 guided wave can effectively reconstruct the shape and size of small and shallow corrosion defects within aluminum plates.

The promoting effect of exercise motivation on physical fitness in college students: a mediation effect model.

BACKGROUND: In recent years, the physical fitness situation of college students is worrying in China. Exercise motivation is of great importance for the physical activity and physical fitness. However, existing studies have paid limited attention to the specific effect between exercise motivation, physical activity and physical fitness, and even less to the different genders and exercise motivation dimensions. This study aimed to investigate the promoting effect of sport motivation on physical fitness with different gender and dimension in college students. METHODS: Physical fitness levels of 2544 college students in Wuhan and Jingzhou city were measured. Exercise motivation and physical activity was assessed using the Chinese version of the motives for physical activities measure-revised scale and the physical activity survey scale, respectively. Correlation analysis and structural equation model were used to explore the relationship between exercise motivation, physical activity and physical fitness. Bootstrap method was used to test the mediating effect. Multilevel regression analysis was used to examine the effects of different dimensions of exercise motivation on PF. RESULTS: The exercise motivation of college students was directly related to physical fitness (effect value: 0.307) or indirectly related through the mediating effect of physical activity (effect value: 0.092). The mediation effect percentage of physical activity on exercise motivation and physical fitness in male (51.20%) was greater than female (27.18%), and the standardized regression coefficient of the health dimension to PF was ß = 0.151, P < 0.001, and the competence dimension to PF was ß = 0.189, P < 0.001. CONCLUSIONS: The exercise motivation of college students can directly influence PF or indirectly influence PF through the mediating effect of PA. The promoting effect of exercise motivation and PA on PF in college students is related to gender and dimensions of exercise motivation. Therefore, we can improve physical fitness levels of college students by promoting their exercise motivation (especially for health motivation and competence motivation) and increasing their participation in physical activity. This study provides new strategies for improving physical fitness in college students.

Comprehensive immunophenotyping of solid tumor-infiltrating immune cells reveals the expression characteristics of LAG-3 and its ligands.

Background: Immune cell expression profiling from patient samples is critical for the successful development of immuno-oncology agents and is useful to understand mechanism-of-action, to identify exploratory biomarkers predictive of response, and to guide treatment selection and combination therapy strategies. LAG-3 is an inhibitory immune checkpoint that can suppress antitumor T-cell responses and targeting LAG-3, in combination with PD-1, is a rational approach to enhance antitumor immunity that has recently demonstrated clinical success. Here, we sought to identify human immune cell subsets that express LAG-3 and its ligands, to characterize the marker expression profile of these subsets, and to investigate the potential relationship between LAG-3 expressing subsets and clinical outcomes to immuno-oncology therapies. Methods: Comprehensive high-parameter immunophenotyping was performed using mass and flow cytometry of tumor-infiltrating lymphocytes (TILs) and peripheral blood mononuclear cells (PBMCs) from two independent cohorts of samples from patients with various solid tumor types. Profiling of circulating immune cells by single cell RNA-seq was conducted on samples from a clinical trial cohort of melanoma patients treated with immunotherapy. Results: LAG-3 was most highly expressed by subsets of tumor-infiltrating CD8 T central memory (TCM) and effector memory (TEM) cells and was frequently co-expressed with PD-1. We determined that these PD-1+ LAG-3+ CD8 memory T cells exhibited a unique marker profile, with greater expression of activation (CD69, HLA-DR), inhibitory (TIM-3, TIGIT, CTLA-4) and stimulatory (4-1BB, ICOS) markers compared to cells that expressed only PD-1 or LAG-3, or that were negative for both checkpoints. In contrast to tumors, LAG-3 expression was more limited in circulating immune cells from healthy donors and solid tumor patients. Additionally, we found abundant expression of the LAG-3 ligands MHC-II and galectin-3 in diverse immune cell types, whereas FGL1 and LSECtin were minimally expressed by immune cells in the tumor microenvironment (TME). Lastly, we found an inverse relationship between baseline and on-treatment levels of circulating LAG3 transcript-expressing CD8 memory T cells and response to combination PD-1 and CTLA-4 blockade in a clinical trial cohort of melanoma patients profiled by scRNAseq. Conclusions: These results provide insights into the nature of LAG-3- and ligand-expressing immune cells within the TME, and suggest a biological basis for informing mechanistic hypotheses, treatment selection strategies, and combination immunotherapy approaches to support continued development of dual PD-1 and LAG-3 blockade.

Ablation of Dual-Specificity Phosphatase 6 Protects against Nonalcoholic Fatty Liver Disease via Cytochrome P450 4A and Mitogen-Activated Protein Kinase.

Dual-specificity phosphatase 6 (DUSP6) is a specific phosphatase for mitogen-activated protein kinase (MAPK). This study used a high-fat diet (HFD)-induced murine nonalcoholic fatty liver disease model to investigate the role of DUSP6 in this disease. Wild-type (WT) and Dusp6-haploinsufficiency mice developed severe obesity and liver pathology consistent with nonalcoholic fatty liver disease when exposed to HFD. In contrast, Dusp6-knockout (KO) mice completely eliminated these phenotypes. Furthermore, primary hepatocytes isolated from WT mice exposed to palmitic and oleic acids exhibited abundant intracellular lipid accumulation, whereas hepatocytes from Dusp6-KO mice showed minimal lipid accumulation. Transcriptome analysis revealed significant down-regulation of genes encoding cytochrome P450 4A (CYP4A), known to promote ω-hydroxylation of fatty acids and hepatic steatosis, in Dusp6-KO hepatocytes compared with that in WT hepatocytes. Diminished CYP4A expression was observed in the liver of Dusp6-KO mice compared with WT and Dusp6-haploinsufficiency mice. Knockdown of DUSP6 in HepG2, a human liver-lineage cell line, also promoted a reduction of lipid accumulation, down-regulation of CYP4A, and up-regulation of phosphorylated/activated MAPK. Furthermore, inhibition of MAPK activity promoted lipid accumulation in DUSP6-knockdown HepG2 cells without affecting CYP4A expression, indicating that CYP4A expression is independent of MAPK activation. These findings highlight the significant role of DUSP6 in HFD-induced steatohepatitis through two distinct pathways involving CYP4A and MAPK.