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11 pages, 798 KiB  
Article
Endophytic Bacteria with Potential Antimicrobial Activity Isolated from Theobroma cacao in Brazilian Amazon
by Lívia Freitas da Silva Pinto, Taynara Cristina Santos Tavares, Oscar Victor Cardenas-Alegria, Elaine Maria Silva Guedes Lobato, Cristina Paiva de Sousa and Adriana Ribeiro Carneiro Nunes
Microorganisms 2025, 13(7), 1686; https://doi.org/10.3390/microorganisms13071686 (registering DOI) - 18 Jul 2025
Abstract
Endophytic bacteria inhabit plant tissues without damaging them and have specialized adaptation capabilities that allow them to establish themselves in this ecological niche. Endophytes produce numerous secondary metabolites with antimicrobial, anticancer, and pesticide properties, among others. In this study, endophytic bacteria were isolated [...] Read more.
Endophytic bacteria inhabit plant tissues without damaging them and have specialized adaptation capabilities that allow them to establish themselves in this ecological niche. Endophytes produce numerous secondary metabolites with antimicrobial, anticancer, and pesticide properties, among others. In this study, endophytic bacteria were isolated and characterized from cocoa plants in a Brazilian municipality, with the view to evaluate their potential antagonistic activity on clinical bacterial strains. The isolates were identified through phenotypic analysis and molecular characterization. After bacterial isolation, it was possible to verify the presence of 11 different endophytic strains, with a bacterial load of up to 6.3 × 103 CFU/g in each plant. The morphological and biochemical profile of the isolates varied. At the taxonomic level, these bacteria showed 99% similarity with the genera Microbacterium, Curtobacterium, Pseudomonas, Bacillus, Ralstonia, and Methylobacterium. The strains of the phylum Actinobacteria, which are known for producing natural bioactive compounds with high biotechnological potential, were effective in inhibiting Staphylococcus aureus ATCC and multidrug-resistant clinical strains. This work aims to expand knowledge about endophytes, with the aim of applying them in other sectors, such as the production of compounds against resistant human pathogens. Full article
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25 pages, 3082 KiB  
Article
Characteristics of Staphylococcus saprophyticus Isolated from Humans and Animals
by Paulina Prorok, Karolina Bierowiec, Milena Skrok, Magdalena Karwańska, Magdalena Siedlecka, Marta Miszczak, Marta Książczyk, Katarzyna Kapczyńska and Krzysztof Rypuła
Int. J. Mol. Sci. 2025, 26(14), 6885; https://doi.org/10.3390/ijms26146885 - 17 Jul 2025
Abstract
Staphylococcus saprophyticus (S. saprophyticus) is an opportunistic coagulase-negative staphylococcus (CoNS) known to cause urinary tract infections in humans and is increasingly recognized in veterinary medicine. The aim of this study was to provide an epidemiological characterization of S. saprophyticus [...] Read more.
Staphylococcus saprophyticus (S. saprophyticus) is an opportunistic coagulase-negative staphylococcus (CoNS) known to cause urinary tract infections in humans and is increasingly recognized in veterinary medicine. The aim of this study was to provide an epidemiological characterization of S. saprophyticus strains and to identify potential virulence factors that may contribute to interspecies transmission. This research is particularly important, as companion animals represent an understudied reservoir of this microorganism, and their role in the spread of resistant pathogens remains insufficiently understood. A total of 61 S. saprophyticus strains isolated from humans, dogs, and cats were analyzed. Identification was performed using MALDI-TOF MS and confirmed by PCR targeting the hrcA gene. Antimicrobial susceptibility was assessed using the disk diffusion and broth microdilution methods, while resistance genes were detected by PCR. The blaZ and mecA genes were present in all strains; additionally, the majority harbored the resistance genes ermA, ermB, tetM, and tetK. Multidrug resistance (MDR) was identified in 21/61 strains (34.4%). Biofilm-forming capacity was temperature-dependent, with the strongest biofilm production observed at 37 °C (70.5%). At 38 °C and 39 °C, the proportion of strong biofilm producers decreased to 50.8% and 52.5%, respectively. All tested strains demonstrated pathogenic potential in the Galleria mellonella larvae infection model, with the highest mortality recorded for selected feline and canine strains. These findings indicate that S. saprophyticus strains from both humans and companion animals possess notable virulence and multidrug resistance. The detection of genotypically and phenotypically resistant strains in animals highlights their potential role as reservoir for zoonotic transmission. Full article
(This article belongs to the Special Issue Molecular Research on Bacteria)
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18 pages, 2644 KiB  
Article
Exploring the Potential of Extracellular Vesicles from Atlantic Cod (Gadus morhua L.) Serum and Mucus for Wound Healing In Vitro
by Stefania D’Alessio, Igor Kraev, Bergljót Magnadóttir and Sigrun Lange
Biology 2025, 14(7), 870; https://doi.org/10.3390/biology14070870 - 17 Jul 2025
Abstract
Novel therapeutic approaches for wound healing have included biomaterials from the Atlantic cod (Gadus morhua L.), with promising results in wound management. The use of extracellular vesicles (EVs), which can be isolated from cod biofluids, remains to be studied. EVs play key [...] Read more.
Novel therapeutic approaches for wound healing have included biomaterials from the Atlantic cod (Gadus morhua L.), with promising results in wound management. The use of extracellular vesicles (EVs), which can be isolated from cod biofluids, remains to be studied. EVs play key roles in cellular communication, and their use both as biomarkers and as therapeutic agents is widely reported in human pathologies, particularly with respect to mesenchymal stem cells. This pilot study characterized the total proteomic cargo content of EVs from cod serum and mucus and assessed the EVs’ potential for regenerative activity in wound-healing processes, using human and mouse fibroblast and keratinocyte in vitro scratch injury models. The pro-regenerative potential of both cod serum EVs and mucus EVs was identified, with differing capacities for accelerating wound closure in fibroblast and keratinocyte cells. This was further supported by varying effects of the cod serum EVs and mucus EVs on cellular vimentin and FGF-2 levels. The serum EV and mucus EV protein cargoes differed with respect to abundance of protein hits and associated enriched functional GO and KEGG pathways, but both were associated with immune, stress and wound-healing processes. Cod EVs may present as innovative therapeutic options for regenerative medicine applications, and our reported findings provide valuable insights for future in-depth studies. Full article
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21 pages, 1088 KiB  
Review
Veterinary Clinics as Reservoirs for Pseudomonas aeruginosa: A Neglected Pathway in One Health Surveillance
by George Cosmin Nadăş, Alice Mathilde Manchon, Cosmina Maria Bouari and Nicodim Iosif Fiț
Antibiotics 2025, 14(7), 720; https://doi.org/10.3390/antibiotics14070720 - 17 Jul 2025
Abstract
Pseudomonas aeruginosa is a highly adaptable opportunistic pathogen with significant clinical relevance in both human and veterinary medicine. Despite its well-documented role in hospital-acquired infections in human healthcare settings, its persistence and transmission within veterinary clinics remain underexplored. This review highlights the overlooked [...] Read more.
Pseudomonas aeruginosa is a highly adaptable opportunistic pathogen with significant clinical relevance in both human and veterinary medicine. Despite its well-documented role in hospital-acquired infections in human healthcare settings, its persistence and transmission within veterinary clinics remain underexplored. This review highlights the overlooked status of veterinary facilities as environmental reservoirs and amplification points for multidrug-resistant (MDR) P. aeruginosa, emphasizing their relevance to One Health surveillance. We examine the bacterium’s environmental survival strategies, including biofilm formation, resistance to disinfectants, and tolerance to nutrient-poor conditions that facilitate the long-term colonization of moist surfaces, drains, medical equipment, and plumbing systems. Common transmission vectors are identified, including asymptomatic animal carriers, contaminated instruments, and the hands of veterinary staff. The review synthesizes current data on antimicrobial resistance in environmental isolates, revealing frequent expression of efflux pumps and mobile resistance genes, and documents the potential for zoonotic transmission to staff and pet owners. Key gaps in environmental monitoring, infection control protocols, and genomic surveillance are identified, with a call for standardized approaches tailored to the veterinary context. Control strategies, including mechanical biofilm disruption, disinfectant cycling, effluent monitoring, and staff hygiene training, are evaluated for feasibility and impact. The article concludes with a One Health framework outlining cross-species and environmental transmission pathways. It advocates for harmonized surveillance, infrastructure improvements, and intersectoral collaboration to reduce the risk posed by MDR P. aeruginosa within veterinary clinical environments and beyond. By addressing these blind spots, veterinary facilities can become proactive partners in antimicrobial stewardship and global resistance mitigation. Full article
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13 pages, 1243 KiB  
Article
A Tandem MS Platform for Simultaneous Determination of Urinary Malondialdehyde and Diphenyl Phosphate
by Gabriela Chango, Diego García-Gómez, Carmelo García Pinto, Encarnación Rodríguez-Gonzalo and José Luis Pérez Pavón
Int. J. Environ. Res. Public Health 2025, 22(7), 1130; https://doi.org/10.3390/ijerph22071130 - 17 Jul 2025
Abstract
This study presents an advanced analytical method for the simultaneous quantification of malondialdehyde (MDA), a biomarker of oxidative stress, and diphenyl phosphate (DPhP), a metabolite of the organophosphate flame retardant triphenyl phosphate (TPhP), in human urine. The method integrates hydrophilic interaction liquid chromatography [...] Read more.
This study presents an advanced analytical method for the simultaneous quantification of malondialdehyde (MDA), a biomarker of oxidative stress, and diphenyl phosphate (DPhP), a metabolite of the organophosphate flame retardant triphenyl phosphate (TPhP), in human urine. The method integrates hydrophilic interaction liquid chromatography (HILIC), a type of liquid chromatography suitable for polar compounds, for MDA separation, and an online restricted access material (RAM), a preconcentration column, for DPhP isolation, achieving high specificity and sensitivity. Validation with certified urine samples confirmed its robustness across diverse analyte concentrations and complex biological matrices. The optimized clean-up steps effectively minimized carryover, allowing for high-throughput analysis. Application to 72 urine samples revealed a significant positive correlation (ρ = 0.702, p-value = 1.9 × 10−7) between MDA and DPhP levels, supporting a potential link between oxidative stress and TPhP exposure. The subset analysis demonstrated a statistically significant moderate positive correlation in women (ρ = 0.622, p-value = 0.020), although this result should be interpreted with caution because of the limited sample size (N = 14). This method provides a powerful tool for biomonitoring oxidative stress and environmental contaminants, offering valuable insights into exposure-related health risks. Full article
(This article belongs to the Special Issue Research on Environmental Exposure, Pollution, and Epidemiology)
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18 pages, 3226 KiB  
Article
Isolation, Identification, and Antibiotic Resistance, CRISPR System Analysis of Escherichia coli from Forest Musk Deer in Western China
by Kaiwei Yang, Xi Wu, Hui Ding, Bingcun Ma, Zengting Li, Yin Wang, Zexiao Yang, Xueping Yao and Yan Luo
Microorganisms 2025, 13(7), 1683; https://doi.org/10.3390/microorganisms13071683 - 17 Jul 2025
Abstract
Escherichia coli (E. coli) is an opportunistic pathogen widely distributed in nature, and multi-drug resistance (MDR) E. coli has been widely recognized as a critical reservoir of resistance genes, posing severe health threats to humans and animals. A total of 288 [...] Read more.
Escherichia coli (E. coli) is an opportunistic pathogen widely distributed in nature, and multi-drug resistance (MDR) E. coli has been widely recognized as a critical reservoir of resistance genes, posing severe health threats to humans and animals. A total of 288 E. coli strains were isolated and purified from fresh fecal samples of forest musk deer collected from farms in Sichuan, Shaanxi, and Yunnan Provinces of China between 2013 and 2023. This study aimed to conduct antibiotic susceptibility testing and resistance gene detection on the isolated forest musk deer-derived E. coli, analyze the correlations between them, investigate the presence of CRISPR systems within the strains, and perform bioinformatics analysis on the CRISPR systems carried by the strains. Results showed that 138 out of 288 E. coli strains were MDR, with the highest resistance to tetracycline (48.3%), cefalexin (45.1%), and doxycycline (41.7%). Prevalent genes were tetA (41.0%), sul2 (30.2%), blaTEM (27.1%), with 29 gene–phenotype pairs correlated. CRISPR system-negative strains had higher resistance rates to 16 antibiotics and lower detection rates only for aac (6′)-Ib-cr, qnrA, and qnrB compared to CRISPR system-positive strains. Regional analysis showed that the problem of drug resistance in Sichuan and Shaanxi was more serious, and that the detection rate of antibiotic resistance genes was relatively high. This study guides E. coli infection control in forest musk deer and enriches resistance research data. Full article
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15 pages, 4191 KiB  
Article
Whole-Genome Sequencing of a Potentially Novel Aeromonas Species Isolated from Diseased Siberian Sturgeon (Acipenser baerii) Using Oxford Nanopore Sequencing
by Akzhigit Mashzhan, Izat Smekenov, Serik Bakiyev, Kalamkas Utegenova, Diana Samatkyzy, Asset Daniyarov, Ulykbek Kairov, Dos Sarbassov and Amangeldy Bissenbaev
Microorganisms 2025, 13(7), 1680; https://doi.org/10.3390/microorganisms13071680 - 17 Jul 2025
Abstract
Aeromonas spp. are opportunistic pathogens that are widely distributed in water sources, with several species being associated with fish and human diseases. We have previously identified an Aeromonas AB005 isolate from diseased Acipencer baerii. This isolate was identified as A. hydrophila based [...] Read more.
Aeromonas spp. are opportunistic pathogens that are widely distributed in water sources, with several species being associated with fish and human diseases. We have previously identified an Aeromonas AB005 isolate from diseased Acipencer baerii. This isolate was identified as A. hydrophila based on the 16S rRNA and gyrB gene sequences. However, this novel strain does not produce indole and tested negative for ornithine decarboxylase and d-xylose fermentation—differences that set it apart from typical A. hydrophila strains. In the present study, this strain was subjected to whole-genome sequencing and compared with the genomes of the type strain (Aeromonas hydrophila ATCC 7966T) and other Aeromonas spp. Comprehensive genome analysis suggests that AB005 represents a distinct species within the genus. The draft genome of the AB005 strain comprises 4,780,815 base pairs with a GC content of 61.2% and contains 6104 predicted protein-coding sequences along with numerous genes implicated in antibiotic resistance. The core/pan-genome analysis reveals extensive genetic diversity, indicative of a dynamic genomic structure. These findings collectively underscore the taxonomic distinction of the AB005 strain as a novel species and highlight its potential pathogenic implications in aquaculture and public health settings. Full article
(This article belongs to the Section Microbial Biotechnology)
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19 pages, 2500 KiB  
Article
Hyperthermia Augments the H1-Histamine Receptor-Mediated Force in the Human Atrium
by Thanh Hoai Pham, Peter Grundig, Britt Hofmann, Uwe Kirchhefer, Joachim Neumann and Ulrich Gergs
Int. J. Mol. Sci. 2025, 26(14), 6842; https://doi.org/10.3390/ijms26146842 (registering DOI) - 16 Jul 2025
Abstract
It was unknown whether hyperthermia increased the efficacy of histamine to raise the force of cardiac contractions via human H1-histamine receptors. To that end, we measured the force in isolated human atrial preparations (HAPs) excised from the right atrium of patients who underwent [...] Read more.
It was unknown whether hyperthermia increased the efficacy of histamine to raise the force of cardiac contractions via human H1-histamine receptors. To that end, we measured the force in isolated human atrial preparations (HAPs) excised from the right atrium of patients who underwent cardiac surgery due to severe two- or three-vessel coronary heart disease. For comparison, we also measured the force in paced (1 Hz) left and spontaneously beating right atrial preparations of transgenic mice overexpressing cardiac human H1-histamine receptors (H1-TG). Histamine (100 µM) was less efficient in raising the force in left atrial preparations from H1 TG mouse atria under hyperthermia than under hypothermia. Oppositely, histamine was more efficient in augmenting force during hyperthermia than during hypothermia in isolated electrically stimulated (1 Hz) HAPs. In sum, the contractile response to activation of H1-histamine receptor in H1-TG mice and in HAPs are opposite with regard to hyperthermia dependence. In patients with fever, histamine might thus be important, to preserve cardiac contractile function as a compensatory mechanism. Full article
(This article belongs to the Section Molecular Pharmacology)
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19 pages, 2160 KiB  
Article
Genetic Diversity and Phylogenetic Analysis Among Multidrug-Resistant Pseudomonas spp. Isolated from Solid Waste Dump Sites and Dairy Farms
by Tuhina Das, Arkaprava Das, Neha Das, Rittika Mukherjee, Mousumi Saha, Dipanwita Das and Agniswar Sarkar
Acta Microbiol. Hell. 2025, 70(3), 30; https://doi.org/10.3390/amh70030030 - 16 Jul 2025
Abstract
The excessive use of antimicrobials drives the emergence of multidrug resistance (MDR) in bacterial strains, which harbor resistance genes to survive under diverse drug pressures. Such resistance can result in life-threatening infections. The predominance of MDR Pseudomonas spp. poses significant challenges to public [...] Read more.
The excessive use of antimicrobials drives the emergence of multidrug resistance (MDR) in bacterial strains, which harbor resistance genes to survive under diverse drug pressures. Such resistance can result in life-threatening infections. The predominance of MDR Pseudomonas spp. poses significant challenges to public health and environmental sustainability, particularly in ecosystems affected by human activities. Characterizing MDR Pseudomonas spp. is crucial for developing effective diagnostic tools and biosecurity protocols, with broader implications for managing other pathogenic bacteria. Strains were diagnosed through 16S rRNA PCR and sequencing, complemented by phylogenetic analysis to evaluate local and global evolutionary connections. Antibiotic susceptibility tests revealed extensive resistance across multiple classes, with MIC values surpassing clinical breakpoints. This study examined the genetic diversity, resistance potential, and phylogenetic relationships among Pseudomonas aeruginosa strain DG2 and Pseudomonas fluorescens strain FM3, which were isolated from solid waste dump sites (n = 30) and dairy farms (n = 22) in West Bengal, India. Phylogenetic analysis reveals distinct clusters that highlight significant geographic linkages and genetic variability among the strains. Significant biofilm production under antibiotic exposure markedly increased resistance levels. RAPD-PCR profiling revealed substantial genetic diversity among the isolates, indicating variations in their genetic makeup. In contrast, SDS-PAGE analysis provided insights into the protein expression patterns that are activated by stress, which are closely linked to MDR. This dual approach offers a clearer perspective on their adaptive responses to environmental stressors. This study underscores the need for vigilant monitoring of MDR Pseudomonas spp. in anthropogenically impacted environments to mitigate risks to human and animal health. Surveillance strategies combining phenotypic and molecular approaches are essential to assess the risks posed by resilient pathogens. Solid waste and dairy farm ecosystems emerge as critical reservoirs for the evolution and dissemination of MDR Pseudomonas spp. Full article
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16 pages, 1988 KiB  
Article
Epidemiological Surveillance, Variability, and Evolution of Isolates Belonging to the Spanish Clone of the 4,[5],12:i:- Monophasic Variant of Salmonella enterica Serovar Typhimurium
by Xenia Vázquez, Patricia García, Javier Fernández, Víctor Ladero, Carlos Rodríguez-Lucas, Jürgen J. Heinisch, Rosaura Rodicio and M. Rosario Rodicio
Antibiotics 2025, 14(7), 711; https://doi.org/10.3390/antibiotics14070711 - 16 Jul 2025
Abstract
Background/Objective: The present study focused on the analysis of the Spanish clone belonging to the successful 4,[5],12:i:- monophasic variant of Salmonella enterica serovar Typhimurium. Methods: All isolates of the clone recovered in a Spanish region from human clinical samples between 2008 and 2018 [...] Read more.
Background/Objective: The present study focused on the analysis of the Spanish clone belonging to the successful 4,[5],12:i:- monophasic variant of Salmonella enterica serovar Typhimurium. Methods: All isolates of the clone recovered in a Spanish region from human clinical samples between 2008 and 2018 (N = 14) were investigated using microbiological approaches and genome sequence analysis. In addition, they were compared with isolates from the years 2000 to 2003 (N = 21), which were previously characterized but had not yet been sequenced. Results: Phylogenetic analyses indicate that all isolates are closely related (differing by 1 to 103 SNPs) but belong to two clades termed A and B. With few exceptions, clade A comprised isolates of the first period, also including two “older” control strains, LSP 389/97 and LSP 272/98. Clade B only contained isolates from the second period. Isolates from both periods were resistant to antibiotics and biocides, with almost all resistance genes located on large IncC plasmids, additionally carrying pSLT-derived virulence genes. The number of resistance genes was highly variable, resulting in a total of 22 ABR (antibiotic biocide resistance) profiles. The number of antibiotic resistance genes, but not that of biocide resistance genes, was considerably lower in isolates from the second than from the first period (with averages of 5.5 versus 9.6 genes). Importantly, IS26, which resides in multiple copies within these plasmids, appears to be playing a crucial role in the evolution of resistance, and it was also responsible for the monophasic phenotype, which was associated with four different deletions eliminating the fljAB region. Conclusions: the observed reduction in the number of antibiotic resistance genes could correlate with the loss of adaptive advantage originating from the ban on the use of antibiotics as feed additives implemented in the European Union since 2006, facilitated by the intrinsic instability of the IncC plasmids. Two consecutive IS26 transposition events, which can explain both the clonal relationship of the isolates and their variability, may account for the observed fljAB deletions. Full article
(This article belongs to the Special Issue Genomic Analysis of Antimicrobial Drug-Resistant Bacteria)
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18 pages, 1422 KiB  
Article
Potable Water Recovery for Space Habitation Systems Using Hybrid Life Support Systems: Biological Pretreatment Coupled with Reverse Osmosis for Humidity Condensate Recovery
by Sunday Adu, William Shane Walker and William Andrew Jackson
Membranes 2025, 15(7), 212; https://doi.org/10.3390/membranes15070212 - 16 Jul 2025
Abstract
The development of efficient and sustainable water recycling systems is essential for long-term human missions and the establishment of space habitats on the Moon, Mars, and beyond. Humidity condensate (HC) is a low-strength wastewater that is currently recycled on the International Space Station [...] Read more.
The development of efficient and sustainable water recycling systems is essential for long-term human missions and the establishment of space habitats on the Moon, Mars, and beyond. Humidity condensate (HC) is a low-strength wastewater that is currently recycled on the International Space Station (ISS). The main contaminants in HC are primarily low-molecular-weight organics and ammonia. This has caused operational issues due to microbial growth in the Water Process Assembly (WPA) storage tank as well as failure of downstream systems. In addition, treatment of this wastewater primarily uses adsorptive and exchange media, which must be continually resupplied and represent a significant life-cycle cost. This study demonstrates the integration of a membrane-aerated biological reactor (MABR) for pretreatment and storage of HC, followed by brackish water reverse osmosis (BWRO). Two system configurations were tested: (1) periodic MABR fluid was sent to batch RO operating at 90% water recovery with the RO concentrate sent to a separate waste tank; and (2) periodic MABR fluid was sent to batch RO operating at 90% recovery with the RO concentrate returned to the MABR (accumulating salinity in the MABR). With an external recycle tank (configuration 2), the system produced 2160 L (i.e., 1080 crew-days) of near potable water (dissolved organic carbon (DOC) < 10 mg/L, total nitrogen (TN) < 12 mg/L, total dissolved solids (TDS) < 30 mg/L) with a single membrane (weight of 260 g). When the MABR was used as the RO recycle tank (configuration 1), 1100 L of permeate could be produced on a single membrane; RO permeate quality was slightly better but generally similar to the first configuration even though no brine was wasted during the run. The results suggest that this hybrid system has the potential to significantly enhance the self-sufficiency of space habitats, supporting sustainable extraterrestrial human habitation, as well as reducing current operational problems on the ISS. These systems may also apply to extreme locations such as remote/isolated terrestrial locations, especially in arid and semi-arid regions. Full article
(This article belongs to the Special Issue Advanced Membranes and Membrane Technologies for Wastewater Treatment)
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19 pages, 827 KiB  
Article
New Derivatives of Caracasine Acid with Anti-Leukemic Activity and Limited Effectiveness in Spheroid Cultures
by Alírica Isabel Suárez, Katiuska Chávez, Jenny Valentina Garmendia, Claudia Valentina De Sanctis, Soňa Gurská, Petr Džubák, Marian Hajduch and Juan Bautista De Sanctis
Pharmaceuticals 2025, 18(7), 1043; https://doi.org/10.3390/ph18071043 - 15 Jul 2025
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Abstract
Background: The natural compounds caracasine acid (1) and its methyl ester, caracasine (2), isolated from the flowers of Croton micans, are effective against several tumor cell lines. Five semi-synthetic derivatives (37) were synthesized based [...] Read more.
Background: The natural compounds caracasine acid (1) and its methyl ester, caracasine (2), isolated from the flowers of Croton micans, are effective against several tumor cell lines. Five semi-synthetic derivatives (37) were synthesized based on these structures. The study aimed to evaluate the cytotoxic activity of these compounds in 2D and spheroid cultures. Methods: The assays were performed in a panel of 12 human cell lines, 8 cancer and 4 normal cell lines. The compounds were evaluated on spheroids derived from the HCT116, HCT116 p53 knockout (p53KO), A549, and U2OS cell lines, as well as mixed spheroids comprising tumor cells and normal fibroblasts. Results: The parent compound (1), the natural ester (2), and two novel derivatives, the anhydride (7) and the cyclohexanol ester (3), demonstrated cytotoxicity against different leukemic cells and HCT116, HCT116 p53 knockout (p53KO), A549, and U2OS cell lines in conventional two-dimensional cultures. Peroxide formation, however, was significantly higher in leukemic cell lines (p < 0.01) in 2D culture as compared with the other tumor cell lines. The compounds did not induce cell death in spheroid cultures; caspases 8, 9, and 3 were not activated upon treatment. Conclusions: These findings indicate potential applications in leukemia treatment, albeit with limited efficacy against solid tumors. Full article
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18 pages, 4537 KiB  
Article
Colostrum-Derived Exosomal Lactoferrin Promotes Skin Fibroblast Regeneration by Suppressing Inflammatory Responses
by Chu-Hsun Cheng, Wei-Jer Hong, Chien-Nien Li, Yung-Hsueh Huang, Jeng-Haw Tsai, Chih-Yuan Huang, Jen-Chin Wu, Chan-Yen Kuo and Wen-Chun Kuo
Curr. Issues Mol. Biol. 2025, 47(7), 549; https://doi.org/10.3390/cimb47070549 - 15 Jul 2025
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Abstract
Lactoferrin (LF), a multifunctional glycoprotein found abundantly in bovine colostrum, is known for its regenerative and anti-inflammatory properties. In this study, we investigated the wound healing and immunomodulatory effects of colostrum-derived exosome-encapsulated lactoferrin (EV-exoLF) on dermal fibroblasts. EV-exoLF was isolated and characterized via [...] Read more.
Lactoferrin (LF), a multifunctional glycoprotein found abundantly in bovine colostrum, is known for its regenerative and anti-inflammatory properties. In this study, we investigated the wound healing and immunomodulatory effects of colostrum-derived exosome-encapsulated lactoferrin (EV-exoLF) on dermal fibroblasts. EV-exoLF was isolated and characterized via nanoparticle tracking analysis and flow cytometry. Functional assays demonstrated that EV-exoLF significantly promoted fibroblast viability and migration in both mouse NIH/3T3 and human HS-68 cell lines. Furthermore, EV-exoLF reduced the expression of pro-inflammatory cytokines (IL-1 and IL-6) and phosphorylated JNK in lipopolysaccharide (LPS)-treated fibroblasts. These findings suggest that EV-exoLF not only enhances fibroblast-mediated wound closure but also mitigates inflammation, highlighting its therapeutic potential in skin regeneration. Colostrum-derived exosomal lactoferrin may serve as a promising natural, cell-free strategy for managing inflammatory skin conditions and improving wound healing outcomes. Full article
(This article belongs to the Special Issue The Role of Bioactives in Inflammation, 2nd Edition)
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15 pages, 3945 KiB  
Article
Modeling Aberrant Angiogenesis in Arteriovenous Malformations Using Endothelial Cells and Organoids for Pharmacological Treatment
by Eun Jung Oh, Hyun Mi Kim, Suin Kwak and Ho Yun Chung
Cells 2025, 14(14), 1081; https://doi.org/10.3390/cells14141081 - 15 Jul 2025
Viewed by 57
Abstract
Arteriovenous malformations (AVMs) are congenital vascular anomalies defined by abnormal direct connections between arteries and veins due to their complex structure or endovascular approaches. Pharmacological strategies targeting the underlying molecular mechanisms are thus gaining increasing attention in an effort to determine the mechanism [...] Read more.
Arteriovenous malformations (AVMs) are congenital vascular anomalies defined by abnormal direct connections between arteries and veins due to their complex structure or endovascular approaches. Pharmacological strategies targeting the underlying molecular mechanisms are thus gaining increasing attention in an effort to determine the mechanism involved in AVM regulation. In this study, we examined 30 human tissue samples, comprising 10 vascular samples, 10 human fibroblasts derived from AVM tissue, and 10 vascular samples derived from healthy individuals. The pharmacological agents thalidomide, U0126, and rapamycin were applied to the isolated endothelial cells (ECs). The pharmacological treatments reduced the proliferation of AVM ECs and downregulated miR-135b-5p, a biomarker associated with AVMs. The expression levels of angiogenesis-related genes, including VEGF, ANG2, FSTL1, and MARCKS, decreased; in comparison, CSPG4, a gene related to capillary networks, was upregulated. Following analysis of these findings, skin samples from 10 AVM patients were reprogrammed into induced pluripotent stem cells (iPSCs) to generate AVM blood vessel organoids. Treatment of these AVM blood vessel organoids with thalidomide, U0126, and rapamycin resulted in a reduction in the expression of the EC markers CD31 and α-SMA. The establishment of AVM blood vessel organoids offers a physiologically relevant in vitro model for disease characterization and drug screening. The authors of future studies should aim to refine this model using advanced techniques, such as microfluidic systems, to more efficiently replicate AVMs’ pathology and support the development of personalized therapies. Full article
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21 pages, 2089 KiB  
Article
Neuropilin-1: A Conserved Entry Receptor for SARS-CoV-2 and a Potential Therapeutic Target
by Vivany Maydel Sierra-Sánchez, Citlali Margarita Blancas-Napoles, Aina Daniela Sánchez-Maldonado, Indira Medina, Rodrigo Romero-Nava, Fengyang Huang, Enrique Hong, Asdrúbal Aguilera-Méndez, Sergio Adrian Ocampo-Ortega and Santiago Villafaña
Biomedicines 2025, 13(7), 1730; https://doi.org/10.3390/biomedicines13071730 - 15 Jul 2025
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Abstract
Background/Objectives: Neuropilin-1 (NRP1) is a key co-receptor for SARS-CoV-2, complementing the ACE2 receptor. Several investigations have documented highly conserved sequences in this receptor, supporting the implication of NRP1 as a key mediator in SARS-CoV-2 cellular entry mechanisms. Methods: To investigate this [...] Read more.
Background/Objectives: Neuropilin-1 (NRP1) is a key co-receptor for SARS-CoV-2, complementing the ACE2 receptor. Several investigations have documented highly conserved sequences in this receptor, supporting the implication of NRP1 as a key mediator in SARS-CoV-2 cellular entry mechanisms. Methods: To investigate this hypothesis, we examined 104,737 SARS-CoV-2 genome fastas from GISAID genomic data, corresponding to isolates collected between 2020 and 2025 in Mexico. Specifically, we focused on the RRAR motif, a known furin-binding site for NRP-1 and the binding site for ACE2 with the spike protein. Our analysis revealed high conservation (>98%) of the RRAR domain compared to a rapidly diminishing ACE2-binding domain. A complementary analysis, using Data from Gene Expression Omnibus (GEO, GSE150316), showed that NRP1 expression in lung tissue remains relatively stable, whereas ACE2 displayed high inter-individual variability and lower abundance compared to NRP1. Based on this evidence, we designed two humans–rats NRP1 siRNAs that were tested in vivo using a melittin-induced lung injury model. Results: The RT-PCR assays confirmed an effective NRP1 knockdown, and the siRNA-treated group showed a significant reduction in the lesions severity. These findings highlight NRP1 as a stable and relevant therapeutic target and suggest the protective potential of siRNA-mediated gene silencing. Conclusions: The evidence presented here supports the rational design of NRP1-directed therapies for multiple circulating SARS-CoV-2 variants in Mexico. Full article
(This article belongs to the Section Cell Biology and Pathology)
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