Search Results (122)

Head and neck squamous cell carcinoma (HNSCC) remains challenging to treat despite multimodal therapeutic approaches. Cisplatin treatment is effective and cost-efficient, although chemoresistance and disease recurrence limit its efficacy. Understanding the mechanisms of cisplatin resistance and the identification of compounds to target resistant tumor cells are critical for improving patient outcomes. We have demonstrated that cisplatin-induced senescent HN30 HNSCC cells can be eliminated by ABT-263 (navitoclax), a BCL-2/BCL-X_L inhibitor that has senolytic properties. Here, we report the development of a cisplatin-resistant cell line (HN30R) for the testing of ABT-263 and the PROTAC BET degraders ARV-825 and ARV-771. ABT-263 was ineffective in sensitizing HN30R cells to cisplatin, largely due to a lack of senescence induction. However, the BET degraders in combination with cisplatin promoted apoptotic cell death in both HN30 and HN30R cells. The effectiveness of ARV-825 did not appear to depend on the cells entering into senescence, indicating that it was not acting as a conventional senolytic. ARV-825 treatment downregulated BRD4 and its downstream targets, c-Myc and Survivin, as well as decreased the expression of RAD51, a DNA repair marker. These results suggest that the BET degraders ARV-825 and ARV-771 may be effective in improving the response of chemoresistant head and neck cancer to cisplatin treatment. Full article

Three-dimensional (3D) cell culture models are widely used to provide a more physiologically relevant microenvironment in which to host and study desired cell types. These models vary in complexity and cost, ranging from simple and inexpensive to highly sophisticated and costly systems. In this study, we introduce a novel translucent multi-compartmentalized stacked multilayered nanocellulose scaffold and describe its fabrication, characterization, and potential application for co-culturing multiple cell types. The scaffold consists of bacterial nanocellulose (BNC) layers separated by interlayers of a lower density of nanocellulose fibers. Using this system, we co-cultured the MDA-MB-231 cell line with two tumor-associated cell types, namely BC-CAFs and M2 macrophages, to simulate the tumor microenvironment (TME). Cells remained viable and metabolically active for up to 15 days. Confocal microscopy showed no signs of cell invasion. However, BC-CAFs and MDA-MB-231 cells were frequently observed within the same layer. The expression of breast cancer-related genes was analyzed to assess the downstream functionality of the cells. We found that the E-cadherin expression was 20% lower in cancer cells co-cultured in the multi-compartmentalized scaffold than in those cultured in 2D plates. Since E-cadherin plays a critical role in preventing the initial dissociation of epithelial cells from the primary tumor mass and is often downregulated in the tumor microenvironment in vivo, this finding suggests that our scaffold more effectively recapitulates the complexity of a tumor microenvironment. Full article

Background/Objectives: Metabolic dysfunction-associated steatotic liver disease (MASLD) is the most prevalent chronic liver disease worldwide, contributing to metabolic dysfunction and increased healthcare costs. The green Mediterranean diet reduces intrahepatic fat and elevates the plasma levels of 2,5-dihydroxybenzoic acid (2,5-DHBA), suggesting a mechanistic role for 2,5-DHBA in hepatic lipid metabolism. This study aimed to evaluate the therapeutic potential of 2,5-DHBA in MASLD and elucidate its molecular mechanism. Methods: Lipid accumulation was assessed in oleic acid-treated HepG2 cells and a high-fat diet (HFD)-induced MASLD mouse model. RNA sequencing, molecular docking, and immunohistochemical staining were performed to investigate the molecular mechanisms, focusing on the chemokine (C-C motif) ligand 2 (CCL2)–CCL2 receptor (CCR2) axis. Results: 2,5-DHBA significantly reduced hepatic lipid accumulation in both HepG2 cells and HFD-fed mice in a dose-dependent manner. RNA sequencing revealed the marked downregulation of CCL2, a key proinflammatory mediator in MASLD pathogenesis. Molecular docking predicted that 2,5-DHBA competed with CCL2 for binding at the CCR2 axis. Immunohistochemistry further confirmed that 2,5-DHBA treatment lowered hepatic CCL2 expression, suppressed nuclear factor-κB activation, and reduced inflammatory cell infiltration. These findings suggest that 2,5-DHBA exerted anti-steatotic effects by modulating the CCL2-CCR2 signaling pathway. Conclusions: This is the first study to demonstrate that 2,5-DHBA attenuates hepatic steatosis via targeting the CCL2-CCR2 axis. These findings highlight its potential as a novel nutraceutical strategy for MASLD treatment. Full article

The diamide insecticide tetraniliprole is a valuable tool for managing major insect pests like the invasive tomato pinworm, Tuta absoluta (Meyrick). However, the mechanisms underlying tetraniliprole resistance, as well as its associated fitness costs, remain unclear. In this study, we assessed the fitness of tetraniliprole-resistant (TetraRS) and susceptible (SS) strains of T. absoluta and conducted Illumina RNA-seq to compare their transcriptomes. We also used nanocarrier-mediated RNA interference (RNAi) to knockdown P450 genes and evaluate their role in tetraniliprole resistance. After eight generations of selection, T. absoluta developed a 20.80-fold resistance to tetraniliprole, accompanied by fitness costs. RNA-seq analysis revealed 3332 differentially expressed genes (DEGs), with 1707 upregulated and 1625 downregulated in the TetraRS compared to the SS strain. Gene Ontology (GO) annotations showed significant enrichment in categories related to metabolic processes, cellular processes, catalytic activity, cellular anatomical entity, and binding. These genes were also identified in key KEGG pathways such as cytochrome P450, drug metabolism, carbon metabolism, oxidative phosphorylation, fatty acid metabolism, and protein processing. RT-qPCR analysis confirmed that P450 genes (CYP405D1, CYP6AB269, and CYP4AU1) were upregulated in TetraRS insects, in line with the RNA-seq results. Cytochrome P450 activity was significantly higher in the TetraRS strain than in the SS strain. Notably, nano-encapsulated dsRNA targeting these overexpressed P450 genes increased the susceptibility of T. absoluta to tetraniliprole. Further, cytochrome P450 activity was significantly reduced following silencing of P450 genes. These findings suggest that multiple genes and pathways, particularly P450 genes, contribute to tetraniliprole resistance in T. absoluta. This study provides valuable insights into the molecular mechanisms underlying insecticide resistance in this key pest species. Full article

Background: Amyotrophic lateral sclerosis (ALS) is a rare, incurable, and fatal neurodegenerative disease that affects the muscles and results in paralysis. The onset and development of ALS involve complex interactions among metabolic signaling, genetic pathways, and external factors (epigenetics). New biomarkers and alternative therapeutic targets have been suggested; nonetheless, the results have been unsatisfactory. Mutations in SOD1, fused in sarcoma (FUS), and TAR DNA-binding protein 43 (TDP-43) have been identified in sporadic amyotrophic lateral sclerosis and approximately 12–20% of familial amyotrophic lateral sclerosis (fALS). Aim: This review analyzes dysregulated microRNA signaling pathways and their interactions with metabolic pathways in the context of ALS progression. Significance: Despite this, biomarkers remain unreliable, and the current medications prolong life without providing a cure. Some proposed approaches to control ALS progression include balancing autophagy and apoptosis, eliminating aggregated proteins, addressing mitochondrial dysfunction, and reducing inflammation. There is a need for studies on new biomarkers, medications, and therapeutic targets. In this context, deregulated circulating microRNAs are attracting attention for new studies on ALS at various phases of the disease. Despite the extensive literature on microRNAs as potential biomarkers for ALS, the proposition for translational clinical use remains limited. Studies have indicated a significant downregulation or upregulation of microRNAs in the motor neurons of ALS patients compared with those with other neurodegenerative disorders and healthy controls. The microRNA biogenesis highlights the importance of this study. MicroRNAs regulate protein synthesis (translation); all human cells express many microRNAs. The complementary structures of microRNA sequences and their mRNA targets allow them to significantly alter cellular and physiological processes. Studies have examined these microRNAs as potential biomarkers for several physiological states and diseases. Comments: The success of these studies may lead to simple, low-cost, and efficient solutions for controlling the progression of ALS and other degenerative diseases. As a result, it is challenging to identify a specific biomarker with total reliability, as a specific microRNA that is increased in one disease phase can decrease in another. These points require careful consideration. They exhibit several complexities and varied interactions, focusing on mRNA targets. The current critical review highlights the potential of microRNAs as biomarkers for diagnosis, prognosis, and therapeutic options in ALS, and raises several points for discussion. Conclusions: The current critical review highlights the potential of microRNAs as biomarkers for diagnosis, prognosis, and therapeutic options in ALS, and raises several points for discussion. Full article

Polyhydroxyalkanoates (PHAs), as biosynthetic and biodegradable polymers, serve as alternatives to petroleum-based plastics, yet face critical cost barriers in large-scale production. While magnetic field (MF) stimulation enhances microbial activity, the optimal MF parameters and metabolic mechanisms for PHA biosynthesis remain unexplored. This study optimized magnetic field parameters to increase PHA biosynthesis in Haloferax mediterranei. A custom-engineered electromagnetic system identified 110 mT of static magnetic field (SMF) as the optimal level for biosynthesis, reaching 77.97 mg/(L·h) PHA volumetric productivity. A pulsed magnetic field caused oxidative stress and impaired substrate uptake despite increasing PHA synthesis. Prolonged SMF exposure (72 h) maximized PHA productivity, while 48 h of exposure attained 90% efficiency. Metabolomics revealed that SMF-driven carbon flux redirection via regulated butanoate metabolism led to a 2.10-fold increase in (R)-3-hydroxybutanoyl-CoA), while downregulating acetoacetate (0.51-fold) and suppressing PHA degradation (0.15-fold). This study pioneers the first application of metabolomics in archaea to decode SMF-induced metabolic rewiring in Haloferax mediterranei. Our findings establish SMF as a scalable bioenhancement tool, offering sustainable solutions for the circular bioeconomy. Full article

Osteoarthritis, affecting over 8 million people in the UK, remains a debilitating condition with limited treatment options. Current therapies primarily address symptoms and can exacerbate joint damage over time. Developing disease-modifying drugs that alleviate inflammation and promote joint regeneration is crucial for long-term patient benefit. This study investigates the potential of exosome-like nano-vesicles isolated from grapefruit juice (GEVs) as a novel therapeutic approach for osteoarthritis. GEVs possess regenerative properties and present a promising avenue for clinical translation. In this study, nano-vesicles were isolated and characterized in terms of protein quantification, size, and morphology. In vitro studies demonstrated the safety and efficacy of GEVs, showing an enhancement in human chondrocyte migratory activity of over 13%. GEVs exhibited a dual mechanism of action, reducing inflammation and oxidative stress while promoting cellular regeneration. Specifically, they reduced the expression of COX2 and PTGS2, markers associated with inflammation and pain sensitization, and enhanced the expression of antioxidant genes SD2 and GPX in osteoarthritic-like chondrocytes. Additionally, GEVs downregulated the expression of ADAMTS-5 and hypertrophic COL10 while upregulating chondrogenic markers ACAN, COL2, and SOX9. This research signifies a significant advancement in osteoarthritis therapy, offering a natural, safe, and cost-effective treatment option with the potential for long-lasting benefits. Clinical translation of GEV therapy holds promise for improving patient outcomes and reducing the burden on healthcare systems. Full article

Chronic wounds (CWs) in humans affect millions of people in the US alone, cost billions of dollars, cause much suffering, and still there are no effective treatments. Patients seek medical care when wound chronicity is already established, making it impossible to investigate factors that initiate chronicity. In this study, we used a diabetic mouse model of CWs that mimics many aspects of chronicity in humans. We performed scRNAseq to compare the cell composition and function during the first 72 h post-injury and profiled 102,737 cells into clusters of all major cell types involved in healing. We found two types of fibroblasts. Fib 1 (pro-healing) was enriched in non-CWs (NCWs) whereas Fib 2 (non-healing) was in CWs. Both showed disrupted proliferation and migration, and extracellular matrix (ECM) deposition in CWs. We identified several subtypes of keratinocytes, all of which were more abundant in NCWs, except for Channel-related keratinocytes, and showed altered migration, apoptosis, and response to oxidative stress (OS) in CWs. Vascular and lymphatic endothelial cells were both less abundant in CWs and both had impaired migration affecting the development of endothelial and lymphatic microvessels. Study of immune cells showed that neutrophils and mast cells are less abundant in CWs and that NCWs contained more proinflammatory macrophages (M1) whereas CWs were enriched in anti-inflammatory macrophages (M2). Also, several genes involved in mitochondrial function were abnormally expressed in CWs, suggesting impaired mitochondrial function and/or higher OS. Heat shock proteins needed for response to OS were downregulated in CWs, potentially leading to higher cellular damage. In conclusion, the initiation of chronicity is multifactorial and involves various cell types and cellular functions, indicating that one type of treatment will not fix all problems, unless the root cause is fundamental to the cell and molecular mechanisms of healing. We propose that such a fundamental process is high OS and its association with wound infection/biofilm. Full article

The South American tomato pinworm, Tuta absoluta (Meyrick), is one of the major invasive pests that causes severe economic damage to several hosts, especially tomato plants, globally. Spinosad, a biopesticide widely used against several insect pests, also shows promising results against T. absoluta. Here, we investigated the evolution of spinosad resistance and its effects on biological traits and related gene effects to fully understand the relationship between resistance degree and associated fitness costs. The spinosad-resistant strain (SpRS) with a moderate level of resistance (14.40-fold) was developed following continuous selection of the susceptible strain (SS) against spinosad for eight generations. Fitness analysis showed that the larval and pupal developmental durations were significantly increased, while the adult emergence was substantially reduced in the SpRS compared to SS. Adult longevity (male and female), fecundity, and hatching rates significantly decreased in the SpRS T. absoluta compared to SS. Additionally, the gene expression analyses indicated the down-regulation of development and reproduction-related genes (Vg, VgR, JHBP, JHAMT, JHE, and JHDK) in the SpRS population. Further, the mRNA expression level of the resistance-related cytochrome P450 gene CYP321C40, followed by CYP4M116, CYP6AW1, CYP339A1, and CYP6AB327, were significantly increased in the SpRS T. absoluta. Taken together, these results indicated that although T. absoluta developed a moderate level of spinosad resistance accompanied by fitness costs, continuous and indiscriminate use might elevate the resistance level. Overall, these findings provide important information about the trade-off between resistance degree and fitness cost that might be crucial to designing resistance management strategies against this key invasive herbivore. Full article

Phenolic compounds are harmful organic pollutants found in wastewater from the chemical and pharmaceutical industries, which are frequently accompanied by high saline concentrations. Microorganism-based biodegradation represents an environmentally friendly and cost-effective strategy for phenol removal. In this study, we isolated a bioflocculant-producing Bacillus paralicheniformis BL-1 that is capable of phenol degradation in high-salinity conditions. Differential gene expression analysis revealed the down-regulation of genes related to the synthesis of extracellular polymeric substances and the up-regulation of poly-γ-glutamate biosynthesis in 10% NaCl conditions. These findings indicate that poly-γ-glutamate is the main large biomolecule produced by B. paralicheniformis BL-1. A further investigation suggested that salinity stress resulted in the down-regulated expression of the genes involved in iron homeostasis. Therefore, alleviating iron limitation by supplying excess iron could improve cell growth and, thus, increase the phenol removal rate and flocculating activity. The productivity of poly-γ-glutamate reached 2.23 g/L, and the phenol removal rate reached 73.83% in the synthetic medium supplemented with 10% NaCl, 500 mg/L phenol, and 250 μM FeCl₃. Full article

Using male sterile (MS) lines instead of normal inbred maternal lines in hybrid seed production can increase the yield and quality with lower production costs. Therefore, developing a new MS germplasm is essential for maize hybrid seed production in the future. Here, we reported a male sterility gene ms*-N125, cloned from a newly found MS mutant ms*-N125. This mutant has an underdeveloped tassel that showed impaired glumes and shriveled anthers without pollen grains. The MS locus of ms*-N125 was mapped precisely to a 112-kb-interval on the chromosome 5. This interval contains only three candidate genes, Zm958, Zm959, and Zm960. Sequencing results showed that only candidate Zm960 harbored a 548-bp transposable element (TE) in its 9th exon, and the two other candidate genes were found to have no genetic variations between the mutant and wild type (WT). Thus, Zm960 is the only candidate gene for male sterility of the mutant ms*-N125. In addition, we screened another recessive MS mutant, ms*-P884, which exhibited similar male sterility phenotypes to ms*-N125. Sequencing Zm960 in ms*-P884 showed a 600-bp TE located in its 2nd exon. Zm960 encodes an ATP-binding cassette in the G subfamily of ABC (ABCG) transporters, ZmABCG2a, with both mutants which harbored an Ac/Ds-like transposon in each. To verify the function of ZmABCG2a for male sterility further, we found an ethyl methanesulfonate (EMS) mutant, zmabcg2a*, which displayed male sterility and tassel phenotypes highly similar to ms*-N125 and ms*-P884, confirming that ZmABCG2a must be the gene for male sterility in maize. In addition, the results of lipid metabolome analysis of ms*-N125 young tassels showed that the total lipid content of the mutant was significantly lower than that of the WT, with 15 subclasses of lipids, including PE (phosphatidylethanolamine), PC (phosphatidylcholine), DG (digalactosyldiacylglycerols), and MGDG (monogalactosyldiacylglycerol) which were significantly down-regulated in the ms*-N125 mutant versus its wild type. In summary, we identified alternate mutations of the ZmABCG2a gene, which may be a potential germplasm for hybrid seed production in maize. Full article

Cultured meat (CM) is derived from the in vitro myogenesis of muscle satellite (stem) cells (MSCs) and offers a promising alternative protein source. However, the development of a cost-effective media formulation that promotes cell growth has yet to be achieved. In this study, laxogenin (LAX) and 5-alpha-hydroxy-laxogenin (5HLAX) were computationally screened against myostatin (MSTN), a negative regulator of muscle mass, because of their antioxidant properties and dual roles as MSTN inhibitors and enhancers of myogenesis regulatory factors. In silico analysis showed LXG and 5HLXG bound to MSTN with binding free energies of −7.90 and −8.50 kcal/mol, respectively. At a concentration of 10 nM, LAX and 5HLAX effectively inhibited the mRNA and protein expressions of MSTN, promoted myogenesis, and enhanced myotube formation and maturation. In addition, by acting as agonists of ROS downregulating factors, they exhibited antioxidative effects. This study shows that supplementation with LAX or 5HLAX at 10 nM in CM production improves texture, quality, and nutritional value. We believe this study fills a research gap on media development for myotube formation and maturation, which are important factors for large-scale in vitro CM production that improve product quality, nutritional value, and efficacy. Full article

Microbial adsorption is a cost-effective and environmentally friendly remediation method for heavy metal pollution. The adsorption mechanism of cadmium (Cd) by bacteria inhabiting extreme environments is largely unexplored. This study describes the biosorption of Cd²⁺ by Sphingomonas sp. M1-B02, which was isolated from the moraine on the north slope of Mount Everest and has a good potential for biosorption. The difference in Cd²⁺ adsorption of the strain after UV irradiation stimulation indicated that the adsorption reached 68.90% in 24 h, but the adsorption after UV irradiation increased to 80.56%. The genome of strain M1-B02 contained antioxidant genes such as mutL, recA, recO, and heavy metal repair genes such as RS14805, apaG, chrA. Hydroxyl, nitro, and etceteras bonds on the bacterial surface were involved in Cd²⁺ adsorption through complexation reactions. The metabolites of the strains were significantly different after 24 h of Cd²⁺ stress, with pyocyanin, L-proline, hypoxanthine, etc., being downregulated and presumably involved in Cd²⁺ biosorption and upregulated after UV-C irradiation, which may explain the increase in Cd²⁺ adsorption capacity of the strain after UV-C irradiation, while the strain improved the metabolism of the antioxidant metabolite carnosine, indirectly increasing the adsorption capacity of the strains for Cd²⁺. Full article

Broad-spectrum crop protection technologies, such as abamectin and bifenthrin, are globally relied upon to curb the existential threats from economic crop pests such as the generalist herbivore Tetranychus urticae Koch (TSSM). However, the rising cost of discovering and registering new acaricides, particularly for specialty crops, along with the increasing risk of pesticide resistance development, underscores the urgent need to preserve the efficacy of currently registered acaricides. This study examined the overall genetic mechanism underlying adaptation to abamectin and bifenthrin in T. urticae populations from commercial hop fields in the Pacific Northwestern region of the USA. A transcriptomic study was conducted using four populations (susceptible, abamectin-resistant, and two bifenthrin-resistant populations). Differential gene expression analysis revealed a notable disparity, with significantly more downregulated genes than upregulated genes in both resistant populations. Gene ontology enrichment analysis revealed a striking consistency among all three resistant populations, with downregulated genes predominately associated with chitin metabolism. In contrast, upregulated genes in the resistant populations were linked to biological processes, such as peptidase activity and oxidoreductase activity. Proteolytic activity by peptidase enzymes in abamectin- and bifenthrin-resistant TSSM populations may suggest their involvement in acaricide metabolism. These findings provide valuable insights into the molecular mechanisms underlying acaricide resistance in the TSSM. This knowledge can be utilized to develop innovative pesticides and molecular diagnostic tools for effectively monitoring and managing resistant TSSM populations. Full article

The red palm weevil (RPW) is an invasive pest that causes devastating damage to a variety of palm plants, which exhibit specific immune priming to Bacillus thuringiensis (Bt). However, immune priming in RPW may incur a high fitness cost, and its molecular signaling pathways have not yet been reported. Here, we investigated the effect of Bt priming on RPW development and subsequently analyzed the hormonal and immune-related molecular pathways influencing the fitness cost induced by Bt priming. Bt priming delayed the body weight gain of fifth-instar larvae and prolonged their developmental duration. Bt priming significantly reduced the 20-hydroxyecdysone (20E) content in RPW hemolymph, and the expression levels of the 20E biosynthesis-related genes SHADOW and SHADE were significantly downregulated. Furthermore, we analyzed Toll pathway genes influencing Bt priming and found that only Spätzle (SPZ) transcription was significantly activated under Bt priming. After silencing SPZ expression, the negative effects of Bt priming on development, SHADOW expression, and 20E synthesis were eliminated, thereby suggesting that SPZ is a key molecular signal mediating developmental and immune trade-offs induced by Bt priming. Our results elucidate the molecular cascade pathway of immune priming and provide new targets for improving the efficiency of RPW biological controls. Full article