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25 pages, 5052 KB  
Article
Comprehensive Analysis of the TaABCB Gene Family and the Role of TaABCB7 in the Phosphate Starvation Response in Wheat
by Guoqing Cui, Haigang Wang, Yanzhen Wang, Xia Liu, Menglin Lei, Huibin Qin, Rui Huang, Juan Lu, Zhixin Mu and Yanming Bai
Biology 2025, 14(11), 1525; https://doi.org/10.3390/biology14111525 - 30 Oct 2025
Abstract
The ABCB subfamily, a subset transporter of the ATP-binding cassette (ABC) superfamily, is vital for various plant life processes, especially in the transport of polar auxin and brassinosteroids. Although ABCB transporters have been characterized in diverse plant species, their specific functions in wheat [...] Read more.
The ABCB subfamily, a subset transporter of the ATP-binding cassette (ABC) superfamily, is vital for various plant life processes, especially in the transport of polar auxin and brassinosteroids. Although ABCB transporters have been characterized in diverse plant species, their specific functions in wheat remain largely unexplored. In this study, we identified 99 TaABCB members in wheat and categorized them into four groups based on their conserved domains and phylogenetic relationships. These members were found to be unevenly distributed across all 21 wheat chromosomes. We conducted a comprehensive genome-wide analysis encompassing gene structure, protein motifs, gene duplication events, collinearity, and cis-acting elements. Transcriptome analysis revealed that different TaABCB members displayed distinct expression patterns under phosphate starvation stress. Notably, we discovered that TaABCB7 might play a role in regulating wheat’s phosphate starvation. Crucially, we pinpointed an elite haplotype, H001, of the candidate gene TaABCB7, which has been progressively selected and employed in wheat breeding improvement programs. Overall, this study enhances our comprehensive understanding of TaABCB members and offers a potential gene resource for molecular marker-assisted selection breeding in wheat. Full article
(This article belongs to the Section Plant Science)
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17 pages, 1610 KB  
Systematic Review
Trap of Social Media Algorithms: A Systematic Review of Research on Filter Bubbles, Echo Chambers, and Their Impact on Youth
by Mukhtar Ahmmad, Khurram Shahzad, Abid Iqbal and Mujahid Latif
Societies 2025, 15(11), 301; https://doi.org/10.3390/soc15110301 - 30 Oct 2025
Abstract
This systematic review synthesizes a decade of peer-reviewed research (2015–2025) examining the interplay of filter bubbles, echo chambers, and algorithmic bias in shaping youth engagement within social media. A total of 30 studies were analyzed, using the PRISMA 2020 framework, encompassing computational audits, [...] Read more.
This systematic review synthesizes a decade of peer-reviewed research (2015–2025) examining the interplay of filter bubbles, echo chambers, and algorithmic bias in shaping youth engagement within social media. A total of 30 studies were analyzed, using the PRISMA 2020 framework, encompassing computational audits, simulation modeling, surveys, ethnographic accounts, and mixed-methods designs across diverse platforms, including Facebook, YouTube, Twitter/X, Instagram, TikTok, and Weibo. Results reveal three consistent patterns: (i) algorithmic systems structurally amplify ideological homogeneity, reinforcing selective exposure and limiting viewpoint diversity; (ii) youth demonstrate partial awareness and adaptive strategies to navigate algorithmic feeds, though their agency is constrained by opaque recommender systems and uneven digital literacy; and (iii) echo chambers not only foster ideological polarization but also serve as spaces for identity reinforcement and cultural belonging. Despite these insights, the evidence base suffers from geographic bias toward Western contexts, limited longitudinal research, methodological fragmentation, and conceptual ambiguity in key definitions. This review highlights the need for integrative, cross-cultural, and youth-centered approaches that bridge empirical evidence with lived experiences. Full article
(This article belongs to the Special Issue Algorithm Awareness: Opportunities, Challenges and Impacts on Society)
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22 pages, 1467 KB  
Article
Reactivity of Curcumin: Theoretical Insight from a Systematic Density Functional Theory-Based Review
by Marcin Molski
Int. J. Mol. Sci. 2025, 26(21), 10374; https://doi.org/10.3390/ijms262110374 - 24 Oct 2025
Viewed by 238
Abstract
A comprehensive analysis of key findings derived from density functional theory (DFT) studies reveals that current theoretical data on curcumin remain incomplete, underscoring the need for further computational investigation to achieve a more thorough understanding of its chemical and biological reactivity. This study [...] Read more.
A comprehensive analysis of key findings derived from density functional theory (DFT) studies reveals that current theoretical data on curcumin remain incomplete, underscoring the need for further computational investigation to achieve a more thorough understanding of its chemical and biological reactivity. This study addresses these gaps through four primary objectives: (i) determination of a complete set of thermodynamic descriptors and elucidation of the multi-step anti-radical mechanisms of the neutral, radical, anionic, and radical–anionic forms of curcumin; (ii) calculation of global chemical reactivity descriptors of curcumin in various solvent environments; (iii) theoretical reproduction of experimentally determined pKa values for all active sites within the molecule; and (iv) examination of the effects of dispersion interactions and solvent polarity on the reactivity descriptors of keto–enol forms of curcumin. The results obtained provide enhanced insight into the molecular behavior of curcumin, facilitating improved predictions of its reactivity under diverse conditions. Moreover, the findings indicate a potential structural modification of the keto form of curcumin, involving the attachment of two 4-hydroxy-3-methoxyphenyl-prop-1-en-2-one moieties to the methylene group. The resulting modeled compound, referred to as di-curcumin, exhibits enhanced chemical reactivity and increased anti-radical potential. Full article
(This article belongs to the Section Physical Chemistry and Chemical Physics)
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18 pages, 5185 KB  
Article
Analysis of the Pollen Morphology and Cluster of Different Pummelo Germplasms
by Dongxing Zhao, Guorui Feng, Zixiang Yang, Guanglin Bi, Hongyuan Wei, Ying Sun, Linyang Chen, Yongzhi Yang, Wanyun Li, Yonghui Li, Chun Li and Hualin Yi
Horticulturae 2025, 11(11), 1277; https://doi.org/10.3390/horticulturae11111277 - 24 Oct 2025
Viewed by 308
Abstract
The pollen morphological characteristics of 16 pummelos and 4 other citrus germplasms from Vietnam, Thailand, and China were observed using scanning electron microscopy (SEM). Observations included equatorial view, polar view, and exine ornamentation. The results showed that the pollen of all tested materials [...] Read more.
The pollen morphological characteristics of 16 pummelos and 4 other citrus germplasms from Vietnam, Thailand, and China were observed using scanning electron microscopy (SEM). Observations included equatorial view, polar view, and exine ornamentation. The results showed that the pollen of all tested materials was monads, prolate, or subprolate. The average polar axis length of the pollen ranged from 29.121 to 37.043 µm, and that of the equatorial axis ranged from 19.861 to 25.911 µm. A t-test revealed that the polar axis of pollen from Chinese pummelo germplasms was significantly longer than that of varieties from Southeast Asia, indicating certain geographical differentiation. The apertures were all colporate type, predominantly with four to five colpi (75% of grains); only four colpi were observed in the remainder (25%). The pollen exine ornamentation of pummelo germplasms was perforated, whereas that of other citrus types was reticulate. Pummelos exhibited a smaller pori diameter (0.264–0.673 µm) and wider distance between pori (0.581–1.118 µm), while other citrus species had larger lumina (1.253–1.684 µm) and narrower muri (0.443–0.664 µm). Principal component analysis (PCA) and cluster analysis were performed based on pollen traits, and two principal component factors were extracted. The pummelo germplasms were divided into two subgroups: sweet pummelo and red pummelo, which demonstrated a correlation among their pollen morphology, flesh color, and flesh flavor. The phenotypic diversity of pollen among different pummelo germplasms may provide a valuable auxiliary reference for the identification and systematic classification of pummelos. Full article
(This article belongs to the Section Genetics, Genomics, Breeding, and Biotechnology (G2B2))
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24 pages, 4193 KB  
Article
Reconfigurable Circularly Polarized Phased Array
by Eduardo S. Silveira, Fúlvio F. Oliveira, Bernardo M. Fabiani, Juner M. Vieira, Daniel B. Ferreira and Daniel C. Nascimento
Electronics 2025, 14(21), 4159; https://doi.org/10.3390/electronics14214159 - 24 Oct 2025
Viewed by 314
Abstract
This paper presents the design, construction, and tests of a polarization-reconfigurable phased array antenna. The proposed array allows the polarization at the main lobe maximum direction to be electronically reconfigured between right-hand (RHCP) and left-hand circular polarization (LHCP). Single-fed microstrip antennas, each with [...] Read more.
This paper presents the design, construction, and tests of a polarization-reconfigurable phased array antenna. The proposed array allows the polarization at the main lobe maximum direction to be electronically reconfigured between right-hand (RHCP) and left-hand circular polarization (LHCP). Single-fed microstrip antennas, each with four tunable varicap diodes, are employed in the phased array to achieve a low axial ratio (AR) at the steering angles. Special attention is given to the microstrip antenna design and varicap modeling, which involves the use of measured data and search algorithms running in an electromagnetic/circuit co-simulation environment. To illustrate the proposed approach, a six-element linear phased array at 2.2 GHz has been built and tested in an anechoic chamber. The experimental results demonstrate an AR below 1 dB in both RHCP and LHCP states over a wide range of steering angles, and even in a multibeam configuration, validating our design method. Full article
(This article belongs to the Section Microwave and Wireless Communications)
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13 pages, 1636 KB  
Article
The Identification of a Sub-Micromolar Peptide-Based Protein Arginine Methyltransferase 1 (PRMT1) Inhibitor from a Plate-Based Screening Assay
by Tina M. Sawatzky, Sarah A. Mann, Jordan Shauna Tucker, Aida A. Bibart, Corey P. Causey and Bryan Knuckley
Biomolecules 2025, 15(11), 1494; https://doi.org/10.3390/biom15111494 - 23 Oct 2025
Viewed by 699
Abstract
Post-translational modifications (PTMs) expand the structural diversity of proteins beyond the standard amino acids, influencing protein-protein interactions. Protein methylation, a prevalent PTM, involves the transfer of methyl groups from S-adenosylmethionine (SAM) to lysine and arginine residues. Arginine methylation is catalyzed by the Protein [...] Read more.
Post-translational modifications (PTMs) expand the structural diversity of proteins beyond the standard amino acids, influencing protein-protein interactions. Protein methylation, a prevalent PTM, involves the transfer of methyl groups from S-adenosylmethionine (SAM) to lysine and arginine residues. Arginine methylation is catalyzed by the Protein Arginine Methyltransferase (PRMT) family to yield mono- and dimethylarginine forms. PRMT1, the isozyme responsible for the majority of asymmetric dimethylation (ADMA) is implicated in various diseases, including cancer. Here, we report the synthesis and screening of a second-generation peptide library to identify novel PRMT1 substrates. The library, based on histone peptides, incorporated varying sequences of amino acids, facilitating substrate specificity studies. Screening identified 7 peptide sequences as exceptional PRMT1 substrates, which were confirmed by kinetic analysis. Consensus sequences revealed key recognition elements for PRMT1 catalysis, suggesting roles for small non-polar side chains and specific residues near the substrate arginine. Furthermore, we developed a peptide-based PRMT1 inhibitor by substituting the substrate arginine with a chloroacetamidine warhead. The inhibitor exhibited sub-micromolar inhibitory potency against PRMT1, surpassing previous peptide-based inhibitors. Our findings contribute to understanding PRMT1 substrate specificity and provide a scaffold for developing potent inhibitors targeting PRMT1 in diseases, including cancer. Full article
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12 pages, 22225 KB  
Article
Soil Organic Carbon Mapping Using Multi-Frequency SAR Data and Machine Learning Algorithms
by Pavan Kumar Bellam, Murali Krishna Gumma, Narayanarao Bhogapurapu and Venkata Reddy Keesara
Land 2025, 14(11), 2105; https://doi.org/10.3390/land14112105 - 23 Oct 2025
Viewed by 286
Abstract
Soil organic carbon (SOC) is a critical component of soil health, influencing soil structure, soil water retention capacity, and nutrient cycling while playing a key role in the global carbon cycle. Accurate SOC estimation over croplands is essential for sustainable land management and [...] Read more.
Soil organic carbon (SOC) is a critical component of soil health, influencing soil structure, soil water retention capacity, and nutrient cycling while playing a key role in the global carbon cycle. Accurate SOC estimation over croplands is essential for sustainable land management and climate change mitigation. This study explores a novel approach to SOC estimation using multi-frequency synthetic aperture radar (SAR) data, specifically Sentinel-1 and ALOS-2/PALSAR-2 imagery, combined with advanced machine learning techniques for cropland SOC estimation. Diverse agricultural practices, with major crop types such as rice (Oryza sativa), finger millet (Eleusine coracana), Niger (Guizotia abyssinica), maize (Zea mays), and vegetable cultivation, characterize the study region. By integrating C-band (Sentinel-1) and L-band (ALOS-2/PALSAR-2) SAR data with key polarimetric features such as the C2 matrix, entropy, and degree of polarization, this study enhances SOC estimation. These parameters help distinguish variations in soil moisture, texture, and mineral composition, reducing their confounding effects on SOC estimation. An ensemble model incorporating Random Forest (RF) and neural networks (NNs) was developed to capture the complex relationships between SAR data and SOC. The NN component effectively models complex non-linear relationships, while the RF model helps prevent overfitting. The proposed model achieved a correlation coefficient (r) of 0.64 and a root mean square error (RMSE) of 0.18, demonstrating its predictive capability. In summary, our results offer an efficient approach for enhanced SOC mapping in diverse agricultural landscapes, with ongoing work targeting challenges in data availability to facilitate large-scale SOC mapping. Full article
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26 pages, 2438 KB  
Review
Exosomes in HPV-Associated Cancers: From Biomarkers to Engineered Therapeutics
by Muharrem Okan Cakir, Melis Selek, Betul Yilmaz, Mustafa Ozdogan and G. Hossein Ashrafi
Cancers 2025, 17(20), 3386; https://doi.org/10.3390/cancers17203386 - 21 Oct 2025
Viewed by 506
Abstract
Background/Objectives: Human papillomavirus (HPV) is the main causative agent of cervical cancer and contributes to a significant proportion of other anogenital and oropharyngeal malignancies. The need for better biomarkers and therapeutic approaches in HPV-associated cancers has drawn attention to exosomes, small extracellular vesicles [...] Read more.
Background/Objectives: Human papillomavirus (HPV) is the main causative agent of cervical cancer and contributes to a significant proportion of other anogenital and oropharyngeal malignancies. The need for better biomarkers and therapeutic approaches in HPV-associated cancers has drawn attention to exosomes, small extracellular vesicles known for their stability, biomolecule transport capabilities, and role in cell-to-cell communication. Methods: This review comprehensively evaluates recent literature on the diagnostic, prognostic, and therapeutic applications of small extracellular vesicles, particularly exosomes, in HPV-related cancers. It analyzes findings on exosomal nucleic acids, proteins, and long non-coding RNAs, as well as engineered exosome-based therapies. Results: Exosomal miRNAs (e.g., miR-204-5p, miR-99a-5p, miR-21), proteins (e.g., glycolytic enzymes, HSP90), and lncRNAs (e.g., HOTAIR, DLEU1) have emerged as promising biomarkers for disease detection and monitoring. Exosomal cargo actively participates in HPV-related tumor progression. For example, miRNAs such as miR-21 and miR-146a modulate immune cell polarization and inflammatory signaling, while lncRNAs like HOTAIR promote oncogenic transcriptional programs. Exosomal proteins including HSP90 and ANXA1 facilitate extracellular matrix remodeling and immune evasion, thereby influencing tumor growth and metastasis. In HPV-positive head and neck and cervical cancers, exosomal cargo reflects HPV status, tumor progression, and treatment response. Therapeutic studies demonstrate the utility of exosomes in vaccine delivery, immune modulation, and drug delivery systems, including the use of PROTACs. However, clinical translation faces barriers including isolation protocol standardization, biomarker validation, and scalable production. Conclusions: Exosomes hold great promise for integration into diagnostic and therapeutic workflows for HPV-related cancers. Future research should focus on resolving standardization issues, validating biomarkers in diverse cohorts, and optimizing engineered exosome platforms for targeted therapy. Full article
(This article belongs to the Collection The Development of Anti-cancer Agents)
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16 pages, 1463 KB  
Review
Macrophages in Autoimmune Liver Diseases: From Immune Homeostasis to Precision-Targeted Therapy
by Tianfu Liu, Yizhe Wang, Yichen Huang, Rui Zhao and Haili Shen
Biomedicines 2025, 13(10), 2520; https://doi.org/10.3390/biomedicines13102520 - 16 Oct 2025
Viewed by 482
Abstract
Autoimmune liver diseases (AILDs) represent a diverse spectrum of chronic inflammatory conditions characterized primarily by compromised hepatic immune tolerance, including autoimmune hepatitis (AIH), primary biliary cholangitis (PBC), and primary sclerosing cholangitis (PSC). Recent evidence positions macrophages as pivotal players in AILDs pathogenesis, attributable [...] Read more.
Autoimmune liver diseases (AILDs) represent a diverse spectrum of chronic inflammatory conditions characterized primarily by compromised hepatic immune tolerance, including autoimmune hepatitis (AIH), primary biliary cholangitis (PBC), and primary sclerosing cholangitis (PSC). Recent evidence positions macrophages as pivotal players in AILDs pathogenesis, attributable to their multifaceted roles in inflammation amplification, immune regulation, and fibrogenesis. In the context of AILDs, macrophages exhibit marked polarization imbalance, increased recruitment of monocytes, and impaired clearance of apoptotic cells. Through complex interactions with T lymphocytes and hepatic stellate cells, macrophages orchestrate a pathological milieu promoting inflammation and fibrosis. Notably, diverse programmed cell death (PCD) modalities—autophagy, necroptosis, pyroptosis, and ferroptosis—not only determine macrophage survival and functional phenotype but also significantly impact cytokine release, phenotypic plasticity, and the trajectory of immunopathological progression. This review synthesizes current understandings of macrophage-driven immunoregulatory mechanisms in AILDs, characterizes the regulatory attributes of various macrophage-related PCD processes, and evaluates their relevance in experimental disease models. Furthermore, we highlight recent advancements in biomarker identification and targeted therapeutic strategies. Comprehensive elucidation of the interplay between macrophage immunological activity and programmed cell death pathways promises to inform novel, personalized therapeutic approaches for patients with AILDs. Full article
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14 pages, 1797 KB  
Article
Novel Discorhabdin Derivatives from Antarctic Sponges of the Genus Latrunculia: Expanding the Chemical Diversity of Polar Marine Natural Products
by Sam Afoullouss, Stine S. H. Olsen, Sydney Morrow, Ezequiel Cruz Rosa, Kaley Geu, Nerida G. Wilson and Bill J. Baker
Mar. Drugs 2025, 23(10), 401; https://doi.org/10.3390/md23100401 - 15 Oct 2025
Viewed by 2628
Abstract
In this study, three Antarctic sponges of the genus Latrunculia were investigated, leading to the isolation of five unreported pyrroloiminoquinone alkaloids along with the known metabolite (+)-debromodiscorhabdin A (3). Three of the new metabolites were brominated, while the other two were [...] Read more.
In this study, three Antarctic sponges of the genus Latrunculia were investigated, leading to the isolation of five unreported pyrroloiminoquinone alkaloids along with the known metabolite (+)-debromodiscorhabdin A (3). Three of the new metabolites were brominated, while the other two were found to have a C-5/C-8 sulfur bridge and a C-2/N-18 bridge. Three of the metabolites were shown to have a phenyl ketone substituent on C-14, not previously reported for discorhabdin derivatives. The cytotoxicity against the A549 cell lines was studied and compounds 14 showed activity of 4.3, 1.8, 1.0, and 23.9 µM, respectively, while no inhibition was found for 5 and 6. Full article
(This article belongs to the Section Structural Studies on Marine Natural Products)
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22 pages, 6649 KB  
Article
Multifunctional Metasurface Based on Cascaded Multilayer Modules
by Tongxing Huang, Shuai Huang, Zhijin Wen, Wei Jiang, Jianxun Wang, Yong Luo and Zeiwei Wu
Nanomaterials 2025, 15(20), 1563; https://doi.org/10.3390/nano15201563 - 14 Oct 2025
Viewed by 336
Abstract
This paper proposes a novel design method for multifunctional modular metasurfaces based on cascaded multilayer modules. Strong electromagnetic coupling between cascaded modules and balanced interface impedance achieved through optimized resonator configurations enable broadband operation. By pairwise cascading of the three modules to maximize [...] Read more.
This paper proposes a novel design method for multifunctional modular metasurfaces based on cascaded multilayer modules. Strong electromagnetic coupling between cascaded modules and balanced interface impedance achieved through optimized resonator configurations enable broadband operation. By pairwise cascading of the three modules to maximize utilization and achieve maximum channel count, the system realizes comprehensive electromagnetic wavefront manipulation across 4 broadband frequency ranges, demonstrating diverse functionalities including orbital angular momentum beam generation, polarization conversion, beam splitting, and radar cross-section reduction with 7 operational channels: two reciprocal co-polarized transmission channels at 14–20.7 GHz, individual reflection channels in +z and −z spaces at 32–38 GHz, two reciprocal cross-polarized transmission channels at 11.9–13.2 GHz, and a reflection channel in −z space at 20–28 GHz, spanning both transmission and reflection modes. The proposed cascading method is accomplished through direct attachment assembly, avoiding air coupling while enabling rapid installation and fast functional switching, providing flexibility for multifunctional electromagnetic wave control applications. Full article
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11 pages, 243 KB  
Article
Analysis of Bound Form Terpenes in Different Agricultural Byproducts
by Duyen Bui, Achyut Adhikari, Witoon Prinyawiwatkul and Zhimin Xu
Molecules 2025, 30(20), 4077; https://doi.org/10.3390/molecules30204077 - 14 Oct 2025
Viewed by 298
Abstract
Traditional sample preparation for terpene analysis includes distillation, solvent extraction, and solid phase extraction and is followed by using gas chromatography with a mass spectrometer (GC-MS) to complete identification and quantification. The preparations rely on the volatility and low polarity of terpenes which [...] Read more.
Traditional sample preparation for terpene analysis includes distillation, solvent extraction, and solid phase extraction and is followed by using gas chromatography with a mass spectrometer (GC-MS) to complete identification and quantification. The preparations rely on the volatility and low polarity of terpenes which exist in free form. However, terpenes in bound form are still largely retained in the extracted residues because, by binding with sugar moiety, they have high polarity and water solubility and low volatility. In this study, distributions and profiles of free and bound form terpenes in different fruit and crop byproducts were evaluated by using different extraction media followed by acid hydrolysis. The acid hydrolysis significantly broke down the binding between terpene and sugar moiety and freed the bound terpene. The concentration of bound terpenes in fruit peel or corn silk was much higher than that of originally existing free terpenes. For example, the terpene concentration in watermelon peel increased from 47.0 to 101 μg/g after hydrolysis. The profile of bound terpenes was also more diverse than that of free terpenes. Among the three extraction media, water, ethanol, and acetone, acetone was the best media to extract bound terpenes with over one and a half times higher total bound terpene extraction yield than ethanol or water extract. The findings of this study explored the bound form terpenes in agricultural products which are usually underexplored in current terpene research. It also demonstrated an effective sample preparation and approach for determining bound terpenes in plants. This study could be an initiating effort and work to assist in exploring rarely mindful bound terpenes in foods and plants. The odorless nature and high stability and water solubility of bound terpenes could provide them a great advantage over free terpenes in various applications requiring neutral scent. Full article
23 pages, 2896 KB  
Protocol
A Practical Guide to Developing and Troubleshooting Patient-Derived “Mini-Gut” Colorectal Organoids for Clinical Research
by Rex Devasahayam Arokia Balaya, Zahra Heydari, Gobinda Sarkar, Estela Mariel Cruz Garcia, Jose M. de Hoyos-Vega, Eugene Krueger, Lauren Helgeson, Alexander Revzin, Alexandra Ros, Akhilesh Pandey and Lisa Boardman
Methods Protoc. 2025, 8(5), 121; https://doi.org/10.3390/mps8050121 - 11 Oct 2025
Viewed by 482
Abstract
Patient-derived organoids (PDOs) have emerged as powerful tools in personalized medicine applicable to both non-malignant conditions and to cancer, where they are increasingly used for personalized drug screening and precision treatment strategies in part due to their ability to replicate tumor heterogeneity. They [...] Read more.
Patient-derived organoids (PDOs) have emerged as powerful tools in personalized medicine applicable to both non-malignant conditions and to cancer, where they are increasingly used for personalized drug screening and precision treatment strategies in part due to their ability to replicate tumor heterogeneity. They also serve as study model systems to understand disease mechanisms, pathways, and the impact of ex vivo exposures. We present a detailed step-by-step protocol for generating organoids from normal crypts, polyps, and tumors, including methods for tissue processing, crypt isolation, culture establishment, and the transition from basolateral to apical-out polarity for co-culture and exposure-based studies. The protocol also includes immunofluorescence staining procedures for cellular characterization and quality control measures. Our standardized approach successfully generates organoids from diverse colorectal tissues with high efficiency and reproducibility. This comprehensive guide addresses common technical challenges and provides troubleshooting strategies to improve success rates across different sample types. We believe that this resource will enhance reproducibility in organoid research and expand their utility in translational applications, particularly for personalized medicine approaches in colorectal cancer. Full article
(This article belongs to the Section Tissue Engineering and Organoids)
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19 pages, 5363 KB  
Article
Human Small Airway Epithelia Reveal Dichloroacetate as a Broad-Spectrum Antiviral Against Respiratory Viruses
by Paula Martínez de Iturrate, Bruno Hernáez, Patricia de los Santos, Yolanda Sierra-Palomares, Alba García-Gómez, Alonso Sánchez-Cruz, Catalina Hernández-Sánchez, Luis Rivas, Margarita del Val and Eduardo Rial
Int. J. Mol. Sci. 2025, 26(20), 9853; https://doi.org/10.3390/ijms26209853 - 10 Oct 2025
Viewed by 412
Abstract
Respiratory viral infections are a major cause of morbidity and mortality worldwide. The COVID-19 pandemic has evidenced the need for broad-spectrum antivirals and improved preclinical models that more accurately recapitulate human respiratory disease. These new strategies should also involve the search for drug [...] Read more.
Respiratory viral infections are a major cause of morbidity and mortality worldwide. The COVID-19 pandemic has evidenced the need for broad-spectrum antivirals and improved preclinical models that more accurately recapitulate human respiratory disease. These new strategies should also involve the search for drug targets in the infected cell that hamper the development of resistance and of potential efficacy against diverse viruses. Since many viruses reprogram cellular metabolism to support viral replication, we performed a comparative analysis of inhibitors targeting the PI3K/AKT/mTOR pathway, central to virus-induced metabolic adaptations, using MRC5 lung fibroblasts and Huh7 hepatoma cells. HCoV-229E infection in MRC5 cells caused the expected shift in the energy metabolism but the inhibitors had markedly different effects on the metabolic profile and antiviral activity in these two cell lines. Dichloroacetate (DCA), a clinically approved inhibitor of aerobic glycolysis, showed antiviral activity against HCoV-229E in MRC5 cells, but not in Huh7 cells, underscoring that the screening model is more critical than previously assumed. We further tested DCA in polarized human small airway epithelial cells cultured in air–liquid interface, a 3D model that mimics the human respiratory tract. DCA reduced the viral progeny of HCoV-229E, SARS-CoV-2, and respiratory syncytial virus by 2–3 orders of magnitude, even when administered after infection was established. Our work reinforces the need for advanced human preclinical screening models to identify antivirals that target host metabolic pathways frequently hijacked by respiratory viruses, and establishes DCA as a proof-of-concept candidate. Full article
(This article belongs to the Special Issue Molecular Mechanisms and Treatments Targeting Respiratory Diseases)
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14 pages, 1332 KB  
Article
Permeabilize, but Choose Wisely: Selective Antibiotic Potentiation Through Outer Membrane Disruption in Pseudomonas aeruginosa
by Marine Novelli and Jean-Michel Brunel
Int. J. Mol. Sci. 2025, 26(20), 9844; https://doi.org/10.3390/ijms26209844 - 10 Oct 2025
Viewed by 459
Abstract
Most clinically used antibiotics exert their effects by targeting essential intracellular components of bacterial cells. Therefore, enhancing their ability to traverse the bacterial envelope is crucial for restoring or improving therapeutic efficacy. We investigated the potential of outer membrane (OM)-disrupting agents—EDTA, NV716, colistin, [...] Read more.
Most clinically used antibiotics exert their effects by targeting essential intracellular components of bacterial cells. Therefore, enhancing their ability to traverse the bacterial envelope is crucial for restoring or improving therapeutic efficacy. We investigated the potential of outer membrane (OM)-disrupting agents—EDTA, NV716, colistin, and squalamine—to potentiate antibiotic activity against the multi-drug-resistant pathogen Pseudomonas aeruginosa. Our objective was to assess the therapeutic value of this strategy while also delineating its limitations by comparing responses across antibiotic classes with diverse chemical structures and pharmacological profiles. Beyond lipophilicity, we analyzed three additional physicochemical descriptors likely to influence OM permeability: molecular surface area, polarizability, and polar surface area. Our findings offer practical insights for the rational design of antibiotic–adjuvant combinations. While each descriptor provides valuable interpretive information, none alone reliably predicts OM-mediated potentiation. Instead, these factors should be viewed collectively within a multidimensional physicochemical profile, where optimal ranges of size, polarity, and lipophilicity act synergistically to enhance antibiotic uptake. By defining a shared multidimensional “responsive zone,” we propose a framework to guide the selection or design of antibiotics compatible with OM-disrupting strategies, potentially enabling the repurposing of antibiotics limited by poor OM permeability. Full article
(This article belongs to the Section Molecular Microbiology)
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