Colorectal cancer (CRC) is one of the most common cancers worldwide and is often treated with chemotherapy. However, systemic toxicity, non-specificity, and drug resistance are major challenges associated with chemotherapeutic drugs. Nanocarriers such as niosomes (NIOs) can enhance drug accumulation at the tumor site while minimizing systemic side effects.

Root rot disease in Panax notoginseng, primarily caused by the pathogenic fungus Fusarium solani, significantly impacts the growth and production of this medicinal herb. To elucidate the defence mechanisms of P. notoginseng against root rot, we employed proteomics and gas chromatography-mass spectrometry (GC-MS)-based metabolomics analyses. These analyses revealed significant accumulations of metabolites involved in phenylpropanoid, terpenoid and steroid biosynthesis pathways in F. solani-infected P. notoginseng roots. This accumulation correlated with the up-regulation of synthetases in these pathways as indicated by proteomics data. Focusing on stigmasterol, a representative steroid with differential accumulation levels, and its biosynthesis gene PnCYP710A, we investigated the role of stigmasterol metabolism in the defence response against root rot. Stigmasterol exhibited significant inhibitory effects on spore germination and hyphal growth of F. solani. Furthermore, PnCYP710A was up-regulated upon F. solani infection and induced by hormonal signals such as methyl jasmonate (MeJA). Overexpression of PnCYP710A in tobacco enhanced resistance to F. solani, up-regulated expression of JA biosynthesis/signalling pathway-related genes, increased accumulation of stigmasterol/lignin/callus, and maintained reactive oxygen species homeostasis during F. solani infection. Conversely, RNA interference (RNAi) of PnCYP710A in P. notoginseng yielded opposite effects. Additionally, PnWRKY4 positively regulated the transcription level of PnCYP710A by binding to its promoter. In summary, this study not only identifies volatile metabolites and proteins involved in the defence response of P. notoginseng against root rot but also discovers that PnWRKY4 activates stigmasterol biosynthesis to resist root rot pathogen infection.

Melatonin (MT) is a growth regulator that influences anthocyanin synthesis during plant growth. However, the regulation mechanism of MT on the coloration of plum peels remains unclear. Here, the effects of MT on the anthocyanin accumulation and coloration in Chinese plum peels were examined after MT (100 μmol/L) or water (control) treatment. The results showed that MT treatment accelerated the changes in plum peels from green to red by increasing the anthocyanin and phenolic contents. MT treatment also increased the antioxidant capacity, including the scavenging capacity of 2,2-diphenyl-1-picrylhydrazyl (DPPH) and ferric reducing antioxidant power (FRAP); and promoted the enzyme activities of anthocyanin synthesis, such as chalcone synthase (CHS), flavanone 3-hydroxylase (F3H), chalcone isomerase (CHI), and UDP-glucose 3-O-glucosyltransferase (UFGT). Moreover, transcriptome analysis indicated that MT treatment up-regulated the expression of several key anthocyanin-related genes, including CHS, F3H, dihydroflavonol 4-reductase (DFR), anthocyanidin synthase (ANS), and UFGT, revealing the regulatory role of MT in anthocyanin metabolism. In conclusion, MT treatment can promote the anthocyanin accumulation and coloration in plum peels.

Alcohol exposure augments the expression and signaling of transforming growth factor-beta (TGFβ), leading to fibroproliferation. We showed that inhibition of TGFβ receptor type 1 (TGFβR1) mitigates the effect of alcohol in the lung. We further demonstrated that alcohol modulates TGFβ signaling, partly through its ability to modify microRNA (miRNA or miR) expressions in the lung. The mechanism by which alcohol mediates miRNA expressions and how its connection to TGFβ signaling has not been well elucidated. Circular RNAs (circRNAs or circ) have emerged as potential therapeutic targets due to their stability, tissue specificity, and ability to modify miRNAs. Human and murine lung fibroblasts (LF) were treated ± ethanol (60 mM). Samples were analyzed for TGFβR1 and circ-RORβ levels. In silico analysis was performed to identify common miRNAs with binding sites for both TGFβR1 and circ-RORβ. The top 10 miRNA levels in human LF (HLF) ± ethanol were analyzed. Human LF were treated with circ-RORβ anti-sense oligonucleotide (ASO) ± ethanol, and samples were collected for miR-140-3p, TGFβR1, fibronectin (FN1), and α-smooth muscle actin (αSMA) levels, and stress fiber formation analyses. In parallel, HLF were treated with mimic miR-140-3p and analyzed for TGFβR1 mRNA levels. HLF were treated ± RORβ silencing RNA, then analyzed for RORβ, circ-RORβ, miR-140-3p, and TGFβR1 levels. We showed that ethanol upregulates TGFβR1 and circ-RORβ in ethanol-treated LF. In silico analysis revealed that miR-140-3p has putative binding sites for both TGFβR1 3' untranslated region (UTR) and circ-RORβ. We demonstrated that ethanol exposure attenuated LF 's miR-140-3p expression, and inhibition of circ-RORβ abrogated ethanol-mediated miR-140-3p suppression. Inhibition of circ-RORβ attenuated ethanol-induced TGFβR1, FN1, and αSMA expressions, as well as αSMA stress fiber formation in LF, while inhibition of RORβ did not alter circ-RORβ, miR-140-3p, or TGFβR1 levels. Overexpression of miR-140-3p promotes TGFβR1 mRNA degradation and inhibits ethanol-induced TGFβR1 expression while has no impact on circ-RORβ level. Taken together, our findings suggest the potential role of circ-RORβ-miR-140-3p-TGFβR1 axis in alcohol-induced TGFβ signaling and fibroblast-to-myofibroblast differentiation.

The ubiquitin system regulates eukaryotic physiology by modifying myriad substrate proteins. Substrate specificity and the assembly of ubiquitin signals are determined by ubiquitin ligases, some of which also modify non-protein biomolecules. Here we expand this substrate realm, revealing that the human ligase HUWE1 can target drug-like small molecules. We demonstrate that compounds previously reported as HUWE1 inhibitors present substrates of their target ligase. Compound ubiquitination is driven by the canonical catalytic cascade, linking ubiquitin to the compound's primary amino group. In vitro, the modification is selectively catalyzed by HUWE1, allowing the compounds to compete with protein substrates. We establish cellular detection methods, confirming HUWE1 promotes - but does not exclusively drive - compound ubiquitination in cells. Converting the existing compounds into specific HUWE1 substrates or inhibitors thus requires enhanced specificity. More broadly, our findings open avenues for harnessing the ubiquitin system to transform exogenous small molecules into novel chemical modalities within cells.

Chemotherapy is the leading treatment for acute lymphoblastic leukemia (ALL). However, many ALL patients eventually develop relapses, the treatment of which remains a major challenge due to their chemoresistance phenotype. As a step towards this end, we here uncovered that relapsed ALL specimens exhibit a significantly lower expression of STAT6 but not of other STATs, when compared with their paired diagnosis specimens. Furthermore, STAT6 plays a distinctive role in chemosensitization of ALL cells to cytarabine (Ara-C), and T-box transcription factor 21 (TBX21) emerged as a plausible intrinsic biomarker of this Ara-C chemosensitization. We demonstrate that STAT6 undergoes SUMOylation on Lys-307 and sentrin/SUMO-specific protease 3 (SENP3)-mediated deSUMOylation in ALL cells. Most importantly, Ara-C specifically induced SENP3 expression and SENP3 knockdown sensitized ALL cells to Ara-C, with an impact equivalent to STAT6 knockout. These findings support the feedback resistance conferred upon ALL cells by Ara-C-induced SENP3 expression. Our findings uncover a novel role for STAT6 in ALL resistance to Ara-C and suggest that its targeted deprivation or pharmacological inhibition specifically sensitizes ALL cells to Ara-C, offering a plausible modality to surmount Ara-C resistance in future ALL treatment.

Somatic embryogenesis receptor kinases (SERKs) are pivotal regulators of plant development and stress adaptation, known to integrate multiple signaling pathways, including brassinosteroid (BR)-mediated responses to coordinately modulate stress-related gene expression. While SERKs participate in biotic/abiotic stress regulation, their roles in heavy metal (HM) stress responses and BR-mediated transcriptional control under HM exposure remain unexplored. Therefore, we systematically identified 20 BraSERK genes in Chinese cabbage (Brassica rapa subsp. Pekinensis), revealing conserved domain architectures and stress-responsive expression patterns. Three BraSERK members (BraSERK9, 12, and 17) exhibited significant induction under Cd/Zn stress. The potential regulatory roles of BR on Chinese cabbage exposed to Cd/Zn stress were investigated in this study by exogenous spraying of 24-epibrassinolide (EBR). The results showed that EBR could significantly increase total chlorophyll content, reduce Zn/Cd translocation, increase antioxidant capacity, and thus attenuate Cd/Zn-induced toxicity. Moreover, EBR dynamically regulated BraSERKs expression in a tissue- and metal-specific manner, potentiating BraSERK12/17 induction in Zn-stressed shoots while suppressing BraSERK9. Our findings suggest that BraSERKs may contribute to BR-mediated HM adaptation, while EBR appears to play a dual protective role through both direct phytoprotection and potential modulation of SERK pathway activity under Cd/Zn co-stress conditions. This study advances our understanding of BR-mediated metal tolerance in crops, offering potential strategies to mitigate HM toxicity through targeted SERK pathway modulation.

Gemcitabine is commonly used in the standard first-line treatment of urothelial carcinoma (UC); however, the emergence of drug resistance significantly limits its clinical benefit. The present study aims to investigate the role of CUB domain-containing protein 1 (CDCP1) in mediating resistance to gemcitabine in UC cells. Gemcitabine-resistant T24 (T24-GR) cells exhibited downregulation of human equilibrative nucleoside transporter 1 and upregulation of cytidine deaminase, key regulators of gemcitabine metabolism, as well as increased CDCP1 expression. Notably, silencing CDCP1 reversed these resistance-associated expression patterns. Mechanistically, T24-GR cells displayed elevated expression of CDCP1 and increased phosphorylation of c-Src and PKCδ, indicating activation of downstream survival signaling. Overexpression of CDCP1 in T24-CD cells activated similar pathways and modulated regulators of gemcitabine metabolism. In contrast, CRISPR/Cas9-mediated knockout of CDCP1 in T24-CDKO cells suppressed c-Src/PKCδ signaling and increased sensitivity to gemcitabine-induced cytotoxicity. Using flow cytometry, we observed that treatment with gemcitabine induced apoptosis in parental T24 cells, as indicated by an increase in the sub-G1 population. In contrast, T24-GR and T24-CD cells showed minimal sub-G1 accumulation, suggesting resistance to gemcitabine-induced apoptosis. Western blot analysis revealed decreased levels of cleaved caspase-3 and cleaved poly(ADP-ribose) polymerase in T24-GR and T24-CD cells following gemcitabine exposure, whereas these markers were upregulated in parental T24 and T24-CDKO cells. Furthermore, the knockdown of CDCP1 and the utilization of c-Src/PKCδ signaling inhibitors in T24-GR cells led to the restoration of sensitivity to gemcitabine. By suppressing apoptosis and altering drug metabolism pathways, highlighting CDCP1 as a potential therapeutic target for overcoming gemcitabine resistance in UC.

Widespread use of Zinc Oxide Nanoparticles (ZnO NPs) raises concerns about potential health risks, particularly following maternal exposure during critical developmental windows. The impact of exposure on offspring brain development remains unclear. The work aims to investigate the neurodevelopmental consequences of maternal ZnO NP exposure during gestation, lactation, or both periods in male rat offspring. Pregnant rats were administered ZnO NPs (< 100 nm) or vehicle. Offspring developmental parameters and brain tissues were analyzed at postnatal day 60. Assessments included oxidative stress markers (8-OHdG, MDA, NO), antioxidant (GSH, GST, GPX, SOD, CAT), cholinergic function (AChE), epigenetic markers (DNA methylation, BDNF promoter methylation, miR-34a, miR-29b), neurodegeneration-associated proteins (Aβ1-42, Tau), survival/inflammatory signaling pathways (p-Akt, PI3K mRNA, ERK, Bcl-2, COX2, IL-1β, TNF-α, IL-2, TGF-β), apoptosis (Caspase-3), BDNF mRNA, and brain histology. Maternal ZnO NP exposure significantly reduced offspring brain weight, body weight, and survival index, particularly following combined gestational and lactational exposure. Exposed offspring brains exhibited increased oxidative stress, depleted antioxidant defenses, impaired AChE activity, global DNA hypomethylation with targeted BDNF promoter hypermethylation (correlating with reduced BDNF mRNA), increased Aβ1-42 and Tau accumulation, suppressed PI3K/p-Akt and ERK survival signaling, elevated pro-inflammatory markers (IL-1β, TNF-α, IL-2, COX2, TGF-β), increased apoptosis (Caspase-3) alongside decreased Bcl-2, and dysregulated miRNA expression (increased miR-34a, decreased miR-29b). Histology confirmed duration-dependent neuronal damage. Maternal ZnO NP exposure induces persistent offspring neurotoxicity via oxidative stress, neuroinflammation, apoptosis, and epigenetic dysregulation. This highlights developmental brain vulnerability and the importance of assessing maternal nanoparticle exposure.

Gastric cancer (GC) is one of the most common and lethal malignancies in developing countries, with particularly high prevalence in China. Circular RNAs (circRNAs) have garnered increasing attention for their roles in disease pathogenesis. While circRNAs can be translated, there have been few investigations into the biological functions of "translatable circRNAs" in the initiation and progression of gastric adenocarcinoma. In this study, we identified a circRNA, circMAP3K13, which inhibits the proliferation and migration of GC cells. CircMAP3K13 was found to encode a previously unreported 26 kDa protein, designated MAP3K13-232aa. Mechanistically, MAP3K13-232aa binds directly to the kinase domain of IKKα and enhances its activity, thereby promoting NF-κB signaling. This activation leads to upregulation of NLRP3 and increased cisplatin-induced pyroptosis in GC cells. Moreover, MAP3K13-232aa enhances pyroptosis and reduces tumorigenicity and metastasis in vivo. Taken together, both circMAP3K13 and its encoded protein MAP3K13-232aa represent potential therapeutic targets in GC.

Traditionally, oxygen supplementation has been more strongly associated with yeast vitality and sugar consumption kinetics than with aroma metabolism. This study investigated the effects of oxygen exposure during the exponential growth phase of yeast cells on the polyfunctional thiol profile of wine. Using quantitative real-time RT-PCR, the expression of five genes related to amino acid and peptide uptake, as well as two genes associated with β-lyase activity, was analyzed. UPLC/MS-MS was employed to quantify thiol precursors and volatile thiols in must and wine. The results revealed that oxygen upregulated the expression of genes involved in amino acid and peptide uptake, including GAP1, OPT1, OPT2, PTR2, DAL5, and FOT3, some of which are known thiol precursor permeases. Overexpression of those genes in oxygenated trials was correlated with a reduction in thiol precursor residues in wines, particularly GSH-3MH, which was depleted before entry into the stationary phase. Despite increased gene expression, the concentration of GSH-4MMP was not affected and remained constant during the growth phase. Additionally, the expression of the gene responsible for thiol-releasing β-lyases in yeasts, IRC7, was positively correlated with 4MMP concentration in wines when influenced by oxygen treatment during the initial 72 h of alcoholic fermentation. However, oxygen supplementation was associated with STR3 downregulation, which may have contributed to the lower 3 MH release, despite the increased precursor intake.

The global production and use of polybrominated diphenyl ethers, including 2,2',4,4'-tetrabromodiphenyl ether (BDE-47), have been substantially curtailed in recent decades. However, BDE-47 remains ubiquitously detectable in environmental matrices and human tissues worldwide. In this study, we investigated whether prenatal exposure to BDE-47 disrupts sperm function and DNA methylation in rat offspring. Pregnant rats were treated with BDE-47 from gestational day 0 to parturition. Sperm count, motility, morphology, mitochondrial membrane potential (MMP), reactive oxygen species (ROS) production, sperm chromatin DNA fragmentation index (DFI), serum testosterone, and histopathology were evaluated across generations. Testicular DNA methyltransferase expression and whole-genome bisulfite sequencing were performed to determine the DNA methylation in the F3 generation. BDE-47 exposure altered anogenital distance (AGD), sperm count, motility, morphology, MMP, ROS production, mean DFI, and %DFI in the F1 generation; AGD, morphology, and ROS production in the F2 generation; and AGD, motility, morphology, MMP, ROS production, mean DFI, %DFI, and testicular DNA methyltransferase expression in the F3 generation. Gene ontology analysis revealed that SYCP2, ASMT, and MSH4 were associated with sex differentiation and reproductive development. Our findings indicate that prenatal exposure to BDE-47 exerts transgenerational epigenetic effects, inducing phenotypic changes in the male reproductive system.

Colorectal cancer (CRC) is a common malignant tumor of the digestive tract with a high incidence rate. Herba Sarcandrae (HS) is an antipyretic and has been reported to have anti-cancer effects. This study explored the impacts of β-sitosterol on the sensitivity of CRC to 5-fluorouracil (5-FU) and oxaliplatin and the stability of TBX20 protein in CRC cells. There were 41 HS active ingredients and 265 corresponding potential targets, and 48 potential targets of HS were enriched in CRC. Then, 206 differentially expressed genes (DEGs) related to TBX20 overexpression were screened based on the TCGA database, some of which were associated with TMN stages of COAD patients. Epimedin C, rutin, and β-sitosterol, which could be combined with TBX20, were screened and validated in CRC cells. Functionally, β-sitosterol could suppress proliferation and induce apoptosis of CRC cells. β-sitosterol could also enhance the sensitivity of CRC to 5-FU and oxaliplatin. In xenograft models, both HS and β-sitosterol treatments inhibited tumor growth and upregulated TBX20 protein expression, with β-sitosterol demonstrating a stronger effect. Mechanistically, β-sitosterol may stabilize TBX20 by inhibiting its ubiquitin-mediated degradation. In conclusion, β-sitosterol, as an anti-tumor active component of HS, prevents CRC cell proliferation, and accelerates apoptosis by upregulating TBX20.

Pediatric high-grade gliomas (pHGG) are highly invasive with poor survival outcomes. Timing of Temozolomide administration has been shown to affect survival of adult patients with glioblastoma. We investigated whether pHGGs express circadian genes rhythmically and whether underlying rhythms affect Temozolomide sensitivity. Circadian gene expression in pediatric gliomas was analyzed using PedcBioPortal. Immunoblotting was used to assess protein levels in patient-derived pHGG lines and in high- and low- grade (pLGG) glioma specimens. Rhythmic gene expression in pHGG lines was measured via qPCR, and Temozolomide efficacy was tested during peak versus trough Bmal1 expression. Patient data revealed significantly different mRNA expression in multiple circadian genes between pHGGs and pLGGs, including higher Bmal1 expression in pHGG specimens and lower Rev-Erbα expression. Significantly higher BMAL1 and CLOCK protein levels and lower REV-ERBα were present in pHGG versus pLGG tissue specimens. Our three pHGG lines displayed rhythmic Bmal1 and Rev-Erbα expression post-synchronization. We found significantly decreased proliferation when Temozolomide was applied during trough versus peak Bmal1 expression. The positive arm of the circadian clock appears upregulated in pHGGs compared to pLGGs. Pediatric HGGs rhythmically express circadian genes and exhibit differential Temozolomide sensitivity based on timing of administration.

The 11-Methoxytabersonine (11-MT), a monoterpenoid-indole alkaloid isolated from the leaves of Melodinus henryi, has shown promising therapeutic potential against triple-negative breast cancer (TNBC). This study aimed to evaluate the anti-tumor efficacy of 11-MT and to elucidate its underlying molecular mechanisms in the context of TNBC. The in vitro anti-cancer effects of 11-MT were assessed using Cell Counting Kit-8 (CCK-8), colony formation assays, and flow cytometry. In vivo efficacy and safety were evaluated in a xenograft mouse model using 7-week-old female BALB/cA-nu nude mice. To explore the potential of 11-MT in inducing ferroptosis, transmission electron microscopy (TEM), reactive oxygen species (ROS) detection, mitochondrial membrane potential (MMP) assays, and analysis of glutathione peroxidase 4 (GPX4) expression were conducted. Mechanistic investigations included lentiviral-mediated knockdown of Yin Yang 1 (YY1) and overexpression of cyclin-dependent kinase-like 1 (CDKL1), as well as co-immunoprecipitation (Co-IP), chromatin immunoprecipitation (ChIP), and a series of rescue experiments to delineate the regulatory effect of 11-MT on GPX4 expression. The 11-MT significantly inhibited TNBC cell proliferation, induced cell cycle arrest, and promoted cell death in vitro. In vivo, treatment with 11-MT significantly suppressed tumor growth in TNBC xenograft models without evident toxicity to major organs. Mechanistic studies revealed that 11-MT primarily triggered ferroptosis through downregulation of GPX4, leading to excessive ROS accumulation independent of intracellular glutathione depletion. Furthermore, 11-MT reduced the expression of transcription factor YY1, suppressing CDKL1 transcription, which in turn led to the downregulation of GPX4. In summary, the results revealed that the 11-MT exerts potent anti-tumor activity against TNBC by inducing ferroptosis via the YY1-CDKL1-GPX4 signaling axis, highlighting its potential as a novel therapeutic agent for the treatment of TNBC.

Diabetic wounds exhibit delayed healing, necessitating innovative treatments. This study investigates the wound healing potential of curcumin-loaded chitosan nanoparticles embedded in a sodium alginate-chitosan hydrogel (H/CS-CUR) through in vitro and in vivo assessments. Surface morphology, swelling capacity, antibacterial activity, and biocompatibility of the hydrogels were evaluated. The H/CS-CUR demonstrated sustained release over time and a remarkable swelling capacity of 2075 % within the first 15 min. The H/CS-CUR exhibited higher antibacterial activity compared to the sodium alginate-chitosan hydrogel (H). The expression levels of IGF-1 and TGF-β1 for H/CS-CUR were higher than those for H, indicating enhanced immune response and improved wound healing. In vivo studies demonstrated that H/CS-CUR-treated group accelerated wound closure in diabetic rats compared to the control group. The H/CS-CUR group showed a significant decrease in myeloperoxidase levels compared to the control group, indicating a significant anti-inflammatory effect. The H/CS-CUR group exhibited higher epidermal thickness, dermal thickness, granulation tissue formation, and vessel density compared to other groups in diabetic rats. Expression levels of IGF-1, TGF-β1, and VEGF were higher in H/CS-CUR group, promoting tissue repair, and angiogenesis. Additionally, decreased IL-6 expression suggested anti-inflammatory effects. These findings suggest that H/CS-CUR hydrogel can be an effective wound dressing for accelerated diabetic wound healing.

Migraine is a primary headache disorder without a definite pathophysiology or satisfactory managing strategies. Recently, migraine is primarily a disorder of brain plasticity coupled with altered Ca2+ dynamics regulating gene activity and protein expression. Further, epigenetics provided new insight into migraine pathogenesis and therapeutic response elucidation. Sodium-glucose co-transporter-2-inhibitors (e.g., empagliflozin (EMPA)), are a class of antihyperglycemic agents that can cross the blood-brain-barrier to maintain glucose homeostasis. EMPA possesses myriad pharmacological actions with potential beneficial effects for migraine management. Thus, the current study aimed at exploring EMPA efficacy and mechanisms for treating migraine headache, emphasizing its role in synaptic plasticity and epigenetic mechanisms. Using an animal model of chronic migraine headache, the effect of oral (PO)/intranasal (IN) (brain-targeted)-EMPA versus Zolmitriptan (ZOL) on serum pain marker; Substance-P and migraine symptoms; pain, and photophobia were assessed biochemically and behaviorally. The influence on synaptic plasticity was evaluated by quantifying immunohistochemically stained synaptophysin in brain tissues and assessing calcium/calmodulin-activated-kinases (CaMKIIa)/ Camp-response-element-binding-protein (CREB)/calcitonin-gene-related-peptide (CGRP) or brain-derived-neurotrophic-factor (BDNF) pathways. Further, epigenetic modulation of migraine key genes was evaluated by determining HDAC6, CALCR, and MiR155-5p Moreover, serum glucose and amylin levels were appraised. Both EMPA-PO/IN significantly decreased serum levels of substance-P and pain symptoms, increased brain serotonin levels, synaptophysin expression, and modulated the CaMKIIa/CREB/CGRP/or BDNF pathway. In addition, EMPA-IN, but not ZOL, down-regulated HDAC6, CALCR, and MiR155-5p expressions. Further, unlike EMPA-IN, ZOL, and EMPA-PO demonstrated significant hypoglycemic effects. In conclusion, EMPA-IN shows potential as a novel therapeutic approach for managing migraine headaches by enhancing synaptic plasticity and modulating altered migraine-related epigenetic mechanisms.

Copper oxide nanoparticles (nCuO) are widely used in electronics, energy storage, biomedicine, and various other fields. However, few studies have investigated their impact on the environment and crop growth.

Klebsiella pneumoniae is a multidrug-resistant pathogen implicated in severe community- and hospital-acquired infections such as bacteremia, urinary tract infections, sepsis, and pneumonia. Biofilm formation, driven by extracellular polymeric substances (EPS), enhances its persistence and resistance to antibiotics. This study evaluated the anti-biofilm, antibacterial, and quorum-quenching activities of a novel α-amylase B. cereus-derived α-amylase against clinical isolates of K. pneumoniae.

Melanoma immunotherapy urgently requires approaches that can accurately predict drug responses to minimize unnecessary treatments. Deep learning models have emerged as powerful tools in this domain due to their robust predictive capabilities. Integrating functional characteristics with expression data from mRNA transcripts shows promise for enhancing prediction accuracy. We developed a deep learning model called AMU (Attention mechanism Model for melanoma immUnotherapy) that incorporates a self-attention mechanism to predict clinical responses to immune checkpoint inhibitors in melanoma patients based on mRNA expression profiles. We evaluated AMU's performance against established machine learning approaches including Support Vector Machine (SVM), Random Forest, AdaBoost, XGBoost, and classical Convolutional Neural Networks (CNN). In the validation set (pre-treatment tissue samples), AMU exhibited outstanding performance, with an AUC of 0.941 and an mAP of 0.960. In the test set (post-treatment tissue samples), its AUC was 0.672, and the mAP was 0.800. Model interpretation revealed that the TNF-TNFRSF1A pathway was a crucial pathway influencing the efficacy of melanoma immunotherapy. Additionally, the expression levels of CD80 and CCR3 were closely correlated with the survival rate (hazard ratios of 0.761 and 0.134, respectively) and the response to immune checkpoint inhibitors in melanoma patients. The deep learning model integrated with the self-attention mechanism has demonstrated strong efficacy in processing mRNA expression data for melanoma immunotherapy response prediction. After rigorous evaluation, including batch effect correction and cross-validation, AMU achieved superior performance compared to traditional machine learning approaches. Beyond prediction accuracy, our model interpretation work identified the TNF-TNFRSF1A pathway as potentially crucial in determining melanoma ICI response, a finding aligned with recent experimental evidence. The embedding architecture's ability to capture meaningful gene-gene relationships, partially consistent with established protein interaction networks, suggests broad potential for representation learning in transcriptomic analysis. While acknowledging the limitations of current sample sizes and the need for prospective validation, this work provides both methodological advances in applying transformer architectures to gene expression data and biological insights into immunotherapy response mechanisms. The integration of robust machine learning approaches with domain-specific biological knowledge represents a promising direction for developing clinically relevant biomarkers in precision oncology.