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22 pages, 9110 KB  
Article
Optimization of Ultrasound-Assisted Extraction Process for Silkworm (Antheraea pernyi) Pupae Protein and Its Impact on Functional and Structural Characteristics of Protein
by Yuanyuan Zeng, Hanyu Guo, Yingying Li, Yinghao Xu, Mengli Zhang, Cancan Luo, Yanan Zang and Ji Luo
Molecules 2025, 30(17), 3580; https://doi.org/10.3390/molecules30173580 (registering DOI) - 1 Sep 2025
Abstract
In this study, the ultrasonic-assisted extraction of silkworm pupae protein (SPP) was optimized using response surface methodology. Subsequently, the effects of ultrasonic treatment on the structural and functional characteristics of SPP were systematically analyzed and verified through Pearson correlation analysis. The results showed [...] Read more.
In this study, the ultrasonic-assisted extraction of silkworm pupae protein (SPP) was optimized using response surface methodology. Subsequently, the effects of ultrasonic treatment on the structural and functional characteristics of SPP were systematically analyzed and verified through Pearson correlation analysis. The results showed that the optimal extraction parameters were an ultrasonic treatment time of 120 min, a power of 115 W, a temperature of 54 °C, pH of 10.5, and the average extraction yield was 68.087%. Compared to the control, ultrasonic treatment significantly improved the functional properties of SPP, including solubility (13.13 g/L), water holding capacity (0.18%), oil holding capacity (0.28%), foaming capacity (55.35%), foam stability (12.71%), emulsification activity (2.15 m2/g), emulsification stability (21.95%), gel water holding capacity (11.5%), gel hardness (1.02 N), and gel elasticity (0.49 mm). In addition, the adsorption ability of SPP for 2-octanone and aldehyde was enhanced after ultrasonic treatment. Furthermore, the absorption intensity and maximum wavelength of the SPP fluorescence spectrum extracted via ultrasonic treatment were enhanced, along with the increased surface hydrophobicity and more stable secondary structure which contributed to promoting the functional properties of SPP, proven by Pearson correlation analysis. This study provides a theoretical basis for the further utilization of SPP in the food industry. Full article
15 pages, 4009 KB  
Article
Towards the Potential of Using Downstream-Separated Solvents as the Pulping Liquor of Upstream Lignocellulose Fractionation for Enhanced Acetone–Butanol–Ethanol Production
by Changsheng Su, Yunxing Gao, Gege Zhang, Hao Wen, Rui Chen, Jiajing Wang, Yujie Li, Mingyuan Sun, Jikang Cao and Di Cai
Fermentation 2025, 11(9), 514; https://doi.org/10.3390/fermentation11090514 (registering DOI) - 1 Sep 2025
Abstract
Developing efficient, clean, and sustainable lignocellulose pretreatment technologies is essential for second-generation biofuel production. In this study, we attempted to use downstream-separated binary acetone-water, n-butanol-water, and ethanol-water solutions as the initial liquor for upstream organosolv pulping, in order to achieve the efficient [...] Read more.
Developing efficient, clean, and sustainable lignocellulose pretreatment technologies is essential for second-generation biofuel production. In this study, we attempted to use downstream-separated binary acetone-water, n-butanol-water, and ethanol-water solutions as the initial liquor for upstream organosolv pulping, in order to achieve the efficient and economic closed-circuit clean fractionation of the lignocelluloses for biological acetone–butanol–ethanol (ABE) production. Parameters, including concentration and temperature of the organosolv pulping, were optimized systematically. Results indicated that the 50 wt% ethanol and 30 wt% acetone aqueous solutions and pulping at 200 °C for 1 h exhibited better corn stover fractionation performances with higher fermentable sugar production. The total monosaccharide recovery (including glucose and xylose) was 50.92% and 50.89%, respectively, in subsequent enzymatic saccharification. While pulping corn stover using n-butanol solution as initial liquor showed higher delignification 86.16% (50 wt% of n-butanol and 200 °C for 1 h), the hydrolysate obtained by the organosolv pulps always exhibited good fermentability. A maximized 15.0 g/L of ABE with 0.36 g/g of yield was obtained in Ethanol-200 °C-50% group, corresponding to 112 g of ABE production from 1 kg of raw corn stover. As expected, the lignin specimens fractionated by closed-circuit organosolv pulping exhibited narrow molecule weight distribution, high purity, and high preservation of active groups, which supports further valorization. This novel strategy tightly bridges the upstream and downstream processes of second-generation ABE production, providing a new route for ‘energy-matter intensive’ and environmentally friendly lignocelluloses biorefineries. Full article
(This article belongs to the Special Issue Bioprocesses for Biomass Valorization in Biorefineries)
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17 pages, 1263 KB  
Article
Serotyping and Antibiotic Resistance Profiles of Salmonella spp. and Listeria monocytogenes Strains Isolated from Pet Food and Feed Samples: A One Health Perspective
by Nikolaos D. Andritsos, Antonia Mataragka, Nikolaos Tzimotoudis, Anastasia-Spyridoula Chatzopoulou, Maria Kotsikori and John Ikonomopoulos
Vet. Sci. 2025, 12(9), 844; https://doi.org/10.3390/vetsci12090844 (registering DOI) - 1 Sep 2025
Abstract
Foodborne pathogenic bacteria, like Salmonella spp. and Listeria monocytogenes, can be detected in the primary food production environment. On the other hand, and in the current context of One Health, antimicrobial resistance (AMR) is gaining increased attention worldwide, as it poses significant [...] Read more.
Foodborne pathogenic bacteria, like Salmonella spp. and Listeria monocytogenes, can be detected in the primary food production environment. On the other hand, and in the current context of One Health, antimicrobial resistance (AMR) is gaining increased attention worldwide, as it poses significant threat to public health. The purpose of this study was to confirm the presence of Salmonella spp. and L. monocytogenes in pet food and feed samples, by means of biochemical and/or serological testing of the microbial isolates, and then to screen for AMR against a panel of selected antibiotics. Serotyping of the isolates with multiplex polymerase chain reaction revealed the presence of three of the most common clinical Salmonella serovars (S. Enteritidis, S. Typhimurium, S. Thompson) and the major epidemiologically important L. monocytogenes serotypes (1/2a, 1/2b, 1/2c, 4b) in 15 and 9 confirmed isolates of the pathogens, respectively. Strains of Salmonella spp. showed resistance to tetracycline (n = 3) and combined AMR to tetracycline with either ampicillin (n = 2) or trimethoprim-sulfamethoxazole (n = 3), without any multidrug resistance (MDR) being recorded whatsoever. AMR in L. monocytogenes was documented in 55.5% of the bacterial strains (n = 5) tested against ciprofloxacin, meropenem, penicillin, trimethoprim-sulfamethoxazole, and tetracycline. Alarmingly, one strain of L. monocytogenes was MDR to the latter five antibiotics and deemed resistant in three antibiotic groups (carbapenems, penicillins, tetracyclines), after exhibiting minimum inhibitory concentrations (MICs) to meropenem (MIC = 4 μg/mL), penicillin (MIC = 4 μg/mL), and tetracycline (MIC = 48 μg/mL). To the best of our knowledge, finding an MDR L. monocytogenes in pet food is something reported for the first time herein. The results presented in this study highlight the presence of important foodborne bacterial pathogens, such as Salmonella spp. and L. monocytogenes, with increased AMR to antibiotics and possible MDR at the primary production and at the farm level, due to the misuse of pharmacological substances used to treat zoonotic diseases, probably resulting in detection of resistant strains of these pathogenic bacteria in animal-originated food products (e.g., meat, milk, eggs). Full article
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22 pages, 2681 KB  
Article
Genome Sequence and Characterization of Bacillus cereus Endophytes Isolated from the Alectra sessiliflora and Their Biotechnological Potential
by Khuthadzo Tshishonga and Mahloro Hope Serepa-Dlamini
Microbiol. Res. 2025, 16(9), 198; https://doi.org/10.3390/microbiolres16090198 - 1 Sep 2025
Abstract
Bacillus cereus AS_3 and Bacillus cereus AS_5 are bacterial endophytes isolated from sterilized leaves of the medical plant Alectra sessiliflora, which were previously identified using 16S rRNA sequencing. Here, we present the whole-genome sequencing and annotation of strains AS_3 and AS_5, the [...] Read more.
Bacillus cereus AS_3 and Bacillus cereus AS_5 are bacterial endophytes isolated from sterilized leaves of the medical plant Alectra sessiliflora, which were previously identified using 16S rRNA sequencing. Here, we present the whole-genome sequencing and annotation of strains AS_3 and AS_5, the first genome report of Bacillus cereus strains from A. sessiliflora. The genome of strain AS_3 has 59 contigs, 5 503 542 bp draft circular chromosome, an N50 of 211,274 bp, and an average G+C content of 35.2%; whereas strain AS_5 has 38 contigs, 5,510,121 bp draft circular chromosome, an N50 of 536,033 bp, and an average G+C content of 35.2%. A total of 5679 protein-coding genes, 62 genes coding for RNAs, and 122 pseudogenes in the strain AS_3 genome were identified by the National Center for Biotechnology Information Prokaryotic Annotation pipeline, whereas a total of 5688 gene protein-coding genes were identified in AS_5, with 60 genes coding for RNAs and 120 pseudogenes. Phenotypic analysis and whole-genome sequencing analysis showed that AS_3 and AS_5 share similar characteristics, including Gram-positive, motile, rod-shaped, and endospore-forming have shown a high sequence similarity with Bacillus cereus, type strain ATCC 14579T. Strains AS_3 and AS_5 had genomic digital DNA–DNA hybridization (dDDH) with the type strain Bacillus cereus ATCC 14579T of 85.8% and 86%, respectively, and average nucleotide identities (ANIs) of 98% and 98.01%, respectively. Phylogenomic analysis confirmed that strains AS_3 and AS_5 share very similar genomic and phenotypic characteristics, and are closely related to the type strain Bacillus cereus type strain ATCC 14579T, supporting their classification within the Bacillus cereus species. A total of 10 secondary metabolite gene clusters, including siderophore type petrobactin, terpene type molybdenum cofactor, non-ribosomal peptide synthetase (NRPS) type bacillibactin, and β-lactone type fengycin, were predicted using AntiSMASH software (version 5.0). Putative genes potentially involved in bioremediation and endophytic lifestyle were identified in the genome analysis. Genome sequencing of Bacillus cereus AS_3 and Bacillus cereus AS_5 has provided genomic information and demonstrated potential biotechnological applications. Full article
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12 pages, 914 KB  
Article
Response of Oats to Fertilisation with Compost and Mineral Nitrogen in a Pot Experiment
by Wacław Jarecki, Joanna Korczyk-Szabó, Milan Macák, Anita Zapałowska, Puchooa Daneshwar and Miroslav Habán
Nitrogen 2025, 6(3), 76; https://doi.org/10.3390/nitrogen6030076 (registering DOI) - 1 Sep 2025
Abstract
Organic fertilisers release nutrients more slowly than mineral fertilisers, which is why combining organic and mineral fertilisation gives good results in crop cultivation. In the conducted pot experiment, the reaction of oats to compost fertilisation with or without additional nitrogen mineral fertilisation was [...] Read more.
Organic fertilisers release nutrients more slowly than mineral fertilisers, which is why combining organic and mineral fertilisation gives good results in crop cultivation. In the conducted pot experiment, the reaction of oats to compost fertilisation with or without additional nitrogen mineral fertilisation was examined. The following treatments were used: A, control (no fertilisation); B, compost (sewage sludge 80% + sawdust 20%); C, compost (garden and park waste 80% + sawdust 20%); D, compost (sewage sludge 40% + garden and park waste 40% + sawdust 20%); E, compost B with nitrogen fertilisation (30 N kg ha−1); F, compost C with nitrogen fertilisation (30 N kg ha−1); and G, compost D with nitrogen fertilisation (30 N kg ha−1). The study results indicated that the composts used had an altering impact on the soil’s chemical composition by the end of the experiment. Overall, the lowest levels of nutrients were recorded in the control group, indicating that the composts increased soil fertility. Oat plants were better nourished (SPAD—soil–plant analysis development) after fertilisation with sewage sludge composts than garden and park waste composts. However, the most favourable results were obtained in the treatments where organic fertilisation (composts) was combined with mineral fertilisation (nitrogen). All fertilisation treatments significantly enhanced plant height and the number of panicles in the pot compared to the control. The highest values for the number of grains in the panicle, thousand-grain weight, grain mass from the pot, and protein content in the grain were observed after applying organic–mineral fertilisation. Therefore, fertilisation with composts, especially composts combined with mineral nitrogen, can be recommended for oat cultivation. Full article
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14 pages, 3809 KB  
Article
Assessment of Population Dynamics and Fishery Exploitation of Narrow-Barred Spanish Mackerel (Scomberomorus commerson) in Iranian Waters
by Seyed Ahmadreza Hashemi, Mastooreh Doustdar, Abdullah Al Kindi and Sachinandan Dutta
Oceans 2025, 6(3), 55; https://doi.org/10.3390/oceans6030055 (registering DOI) - 31 Aug 2025
Abstract
The population dynamics and exploitation ratios of the narrow-barred Spanish mackerel (Scomberomorus commerson, Lacépède, 1800) were investigated from August 2020 to February 2023, with data collected from three landing sites (Bushehr, Bandar Abbas, and Chabahar) of Iran waters. During the study [...] Read more.
The population dynamics and exploitation ratios of the narrow-barred Spanish mackerel (Scomberomorus commerson, Lacépède, 1800) were investigated from August 2020 to February 2023, with data collected from three landing sites (Bushehr, Bandar Abbas, and Chabahar) of Iran waters. During the study period, only length was measured for 6504 specimens and both the length and weight were measured for 504 specimens. The mean fork length of the samples was 86 ± 20 cm, and the mean weight was 6230 ± 3742 g. The relationship between length and weight for the total samples was described by the equation W = 0.022 × CL2.76 (n = 504, R2 = 0.90, 95% C.I. for b = 2.52–2.91). The population dynamics indices for S. commerson were as follows: infinite length (Linf) = 173 cm, natural mortality (M) = 0.47 per year, growth coefficient (K) = 0.52 per year, total mortality (Z) = 1.42 ± 0.06 (95% C.I. = 1.36–1.48), fishing mortality (F) = 0.95 per year, and exploitation coefficient (E) = 0.67. The exploitation rate (U) and total stock at the beginning of the year (B0) were 0.6 and 48,333 tons, respectively. The annual average standing stock (Bt) was estimated at 30,526 tons. The exploitation ratio for maximum sustainable yield (EMSY) was 0.50, and fishing mortality at maximum sustainable yield (FMSY) was 1.5. The estimated range for maximum sustainable yield (MSY, in 1000 tons), the B/BMSY ratio, F/FMSY ratio, and saturation (S) ratio of S. commerson in the Iranian part of the Persian Gulf and the Sea of Oman was 20 (17–25), 1.55 (1.25–1.73), 0.90 (0.8–1.12), and 0.45, respectively. The stock of S. commerson is approaching overfishing in Iran waters, imposing immediate management actions to reduce catch and fishing effort. Full article
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21 pages, 5417 KB  
Article
Implementation of a Particle Swarm Optimization Algorithm with a Hooke’s Potential, to Obtain Cluster Structures of Carbon Atoms, and of Tungsten and Oxygen in the Ground State
by Jesús Núñez, Gustavo Liendo-Polanco, Jesús Lezama, Diego Venegas-Yazigi, José Rengel, Ulises Guevara, Pablo Díaz, Eduardo Cisternas, Tamara González-Vega, Laura M. Pérez and David Laroze
Inorganics 2025, 13(9), 293; https://doi.org/10.3390/inorganics13090293 - 31 Aug 2025
Abstract
Particle Swarm Optimization (PSO) is a metaheuristic optimization technique based on population behavior, inspired by the movement of a flock of birds or a school of fish. In this method, particles move in a search space to find the global minimum of an [...] Read more.
Particle Swarm Optimization (PSO) is a metaheuristic optimization technique based on population behavior, inspired by the movement of a flock of birds or a school of fish. In this method, particles move in a search space to find the global minimum of an objective function. In this work, a modified PSO algorithm written in Fortran 90 is proposed. The optimized structures obtained with this algorithm are compared with those obtained using the basin-hopping (BH) method written in Python (3.10), and complemented with density functional theory (DFT) calculations using the Gaussian 09 software. Additionally, the results are compared with the structural parameters reported from single crystal X-ray diffraction data for carbon clusters Cn(n = 3–5), and tungsten–oxygen clusters, WOnm(n = 4–6, m=2,4,6). The PSO algorithm performs the search for the minimum energy of a harmonic potential function in a hyperdimensional space R3N (where N is the number of atoms in the system), updating the global best position ( gbest) and local best position ( pbest), as well as the velocity and position vectors for each swarm cluster. A good approximation of the optimized structures and energies of these clusters was obtained, compared to the geometric optimization and single-point electronic energies calculated with the BH and DFT methods in the Gaussian 09 software. These results suggest that the PSO method, due to its low computational cost, could be useful for approximating a molecular structure associated with the global minimum of potential energy, accelerating the prediction of the most stable configuration or conformation, prior to ab initio electronic structure calculation. Full article
(This article belongs to the Special Issue Optical and Quantum Electronics: Physics and Materials)
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14 pages, 3455 KB  
Article
Three-Dimensional-Printed Polymer–Polymer Composite Electrolytes for All-Solid-State Li Metal Batteries
by Hao Wang, Xin Xiong, Huie Hu and Sijie Liu
Polymers 2025, 17(17), 2369; https://doi.org/10.3390/polym17172369 - 30 Aug 2025
Abstract
High-performance batteries for military and extreme environment applications require alternatives to conventional liquid lithium-ion batteries (LIBs), which suffer from poor low-temperature performance and safety risks. All-solid-state lithium batteries (ASSLBs) offer enhanced safety and superior low-temperature capability. In this work, we designed and fabricated [...] Read more.
High-performance batteries for military and extreme environment applications require alternatives to conventional liquid lithium-ion batteries (LIBs), which suffer from poor low-temperature performance and safety risks. All-solid-state lithium batteries (ASSLBs) offer enhanced safety and superior low-temperature capability. In this work, we designed and fabricated composite solid-state electrolytes using polyvinylidene fluoride (PVDF) and polyacrylic acid (PAA) as polymer matrices, N,N-dimethylformamide (DMF) as the solvent, and lithium bis(trifluoromethane sulfonimide) (LiTFSI) as the lithium salt. Composite solutions with varying PAA mass ratios were prepared. Advanced three-dimensional (3D) printing technology enabled the rapid and precise fabrication of electrolyte membranes. An ionic conductivity of about 2.71 × 10−4 S cm−1 at 25 °C, high mechanical strength, and good thermal properties can be achieved through component and 3D printing process optimization. Assembled LiCoO2||PVDF@PAA||Li ASSLBs delivered an initial discharge capacity of 165.3 mAh/g at 0.1 mA cm−2 (room temperature), maintaining 98% capacity retention after 300 cycles. At 0 °C, these cells provided 157.4 mAh/g initial capacity with 85% retention over 100 cycles at 0.1 mA cm−2. This work identifies the optimal PAA ratio for enhanced electrochemical performance and demonstrates the viability of 3D printing for advanced ASSLB manufacturing. Full article
(This article belongs to the Special Issue Advances in Polymeric Additive Manufacturing—2nd Edition)
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30 pages, 5850 KB  
Article
Integrated Approach for Biochemical and Functional Characterization of Six Clinical Variants of Glucose-6-Phosphate Dehydrogenase
by Beatriz Hernández-Ochoa, Mónica Guadalupe Gualos-González, Jhuremy Alexandra Moreno-Hernández, Laura Morales-Luna, Montserrat Vázquez-Bautista, Luis Miguel Canseco-Ávila, Verónica Pérez de la Cruz, Roberto Arreguin-Espinosa, Elizabeth Hernández-Urzua, Sergio Enríquez-Flores, Ignacio De la Mora-De la Mora, Noemí Cárdenas-Rodríguez, Cindy Bandala, Lucia De Franceschi, Abraham Vidal-Limon and Saúl Gómez-Manzo
Int. J. Mol. Sci. 2025, 26(17), 8464; https://doi.org/10.3390/ijms26178464 (registering DOI) - 30 Aug 2025
Viewed by 47
Abstract
Glucose-6-phosphate dehydrogenase (G6PD) deficiency is a widespread enzymopathy affecting approximately 500 million individuals that represents a significant global health issue. Among the more than 230 identified mutations in the G6PD gene, six class A variants—G6PD Utrecht (Pro409Ser), G6PD Suwalki (Pro409Arg), G6PD Merlo (Pro409Gln), [...] Read more.
Glucose-6-phosphate dehydrogenase (G6PD) deficiency is a widespread enzymopathy affecting approximately 500 million individuals that represents a significant global health issue. Among the more than 230 identified mutations in the G6PD gene, six class A variants—G6PD Utrecht (Pro409Ser), G6PD Suwalki (Pro409Arg), G6PD Merlo (Pro409Gln), G6PD Kawasaki (Gly410Ala), G6PD Shinagawa (Gly410Asp), and G6PD Riverside (Gly410Cys)—are located in the beta-loop near the NADP+ binding site. These mutations are of particular interest due to their association with severe hematologic phenotypes, including chronic hemolytic anemia, as well as their proposed role in the allosteric regulation of G6PD multimerization. This study presents a comprehensive biochemical and functional characterization of these clinically relevant G6PD variants. The variant enzymes were cloned, expressed, and purified for characterization. Kinetic parameters and thermal stability assays, complemented by molecular dynamics simulations (MDS), were employed to elucidate the structural impacts of the mutations. Our results demonstrate that these mutations significantly impair protein function, characterized by reduced affinity for glucose-6-phosphate (G6P) and NADP+, as well as altered thermal stability compared with wild-type G6PD. MDS revealed that point mutations in the βN- and βM-sheets in the NADP+s region propagate subtle conformational changes, ultimately affecting the NADP+c region and the G6P binding cavity. Furthermore, secondary structure element analyses of the simulation data showed that Pro409 and Gly410 point mutations propagate several changes around residues 195–210 (G6P binding site) and 380–400 (NADP+s), explaining their effect on overall catalytic performance. These findings enhance our understanding of the molecular mechanisms underlying G6PD deficiency and its clinical implications, providing a foundation for future therapeutic strategies aimed at mitigating the effects of these variants. Full article
(This article belongs to the Special Issue The Application of Machine Learning to Molecular Dynamics Simulations)
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32 pages, 2911 KB  
Review
Selective Deoxygenation of Biomass Polyols into Diols
by Juan Carlos Serrano-Ruiz
Molecules 2025, 30(17), 3559; https://doi.org/10.3390/molecules30173559 - 30 Aug 2025
Viewed by 47
Abstract
The transition to a sustainable chemical industry necessitates efficient valorization of biomass, with polyols serving as versatile, renewable feedstocks. This comprehensive review, focusing on advancements within the last five years, critically analyzes the selective hydrogenolysis of key biomass-derived polyols—including glycerol, erythritol, xylitol, and [...] Read more.
The transition to a sustainable chemical industry necessitates efficient valorization of biomass, with polyols serving as versatile, renewable feedstocks. This comprehensive review, focusing on advancements within the last five years, critically analyzes the selective hydrogenolysis of key biomass-derived polyols—including glycerol, erythritol, xylitol, and sorbitol—into valuable diols. Emphasis is placed on the intricate catalytic strategies developed to control C–O bond cleavage, preventing undesired C–C scission and cyclization. The review highlights the design of bifunctional catalysts, often integrating noble metals (e.g., Pt, Ru, Ir) with oxophilic promoters (e.g., Re, W, Sn) on tailored supports (e.g., TiO2, Nb2O5, N-doped carbon), which have led to significant improvements in selectivity towards specific diols such as 1,2-propanediol (1,2-PD), 1,3-propanediol (1,3-PD), and ethylene glycol (EG). While substantial progress in mechanistic understanding and catalyst performance has been achieved, challenges persist regarding catalyst stability under harsh hydrothermal conditions, the economic viability of noble metal systems, and the processing of complex polyol mixtures from lignocellulosic hydrolysates. Future directions for this field underscore the imperative for more robust, cost-effective catalysts, advanced computational tools, and intensified process designs to facilitate industrial-scale production of bio-based diols. Full article
(This article belongs to the Section Materials Chemistry)
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20 pages, 2100 KB  
Article
Mutational Analysis Reveals Functional Roles of METTL16 Domains and Residues
by Kurtis Breger, Ian P. Schowe, Noah A. Springer, Nathan J. O’Leary, Agnieszka Ruszkowska, Carlos Resende and Jessica A. Brown
Biology 2025, 14(9), 1145; https://doi.org/10.3390/biology14091145 - 29 Aug 2025
Viewed by 111
Abstract
Human methyltransferase-like protein 16 (METTL16) installs N6-methyladenosine on U6 small nuclear RNA (snRNA) and other RNAs. Multiple X-ray crystal structures of METTL16 have been published; however, we do not yet fully understand the structure–function relationships of specific residues. We designed 38 [...] Read more.
Human methyltransferase-like protein 16 (METTL16) installs N6-methyladenosine on U6 small nuclear RNA (snRNA) and other RNAs. Multiple X-ray crystal structures of METTL16 have been published; however, we do not yet fully understand the structure–function relationships of specific residues. We designed 38 mutants, including seven cancer-associated mutants, and used electrophoretic mobility shift assays and single-turnover kinetic assays to better understand the functional roles of specific domains and amino acid residues in binding to U6 snRNA, formation of the METTL16•U6 snRNA•S-adenosylmethionine (SAM) complex, and the rate of methylation. While point mutations in the methyltransferase domain mildly weaken the binding of METTL16 to U6 snRNA, the C-terminal vertebrate conserved regions (VCRs), particularly the arginine-rich region (R382 to R388), mediate cooperative binding and contribute more to RNA binding. All METTL16 K-loop mutants displayed tighter SAM binding, suggesting that the K-loop blocks SAM binding. In addition, residues E133 and F227 are critical for stabilizing SAM binding. Mutations in the 184NPPF187 catalytic core and R282A abolished methyltransferase activity. Two METTL16 somatic cancer-associated mutants (G110C and R241Dfs*2) displayed reduced methylation activity. This mutational analysis expands our understanding of how specific domains and residues contribute to substrate-binding activity and methylation of U6 snRNA catalyzed by METTL16. Full article
(This article belongs to the Section Biochemistry and Molecular Biology)
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20 pages, 1436 KB  
Article
Thienyl-Based Amides of M2 and Neuraminidase Inhibitors: Synthesis, Structural Characterization, and In Vitro Antiviral Activity Against Influenza A Viruses
by Maya Chochkova, Boyka Stoykova, Magdalena Angelova, Hristina Sbirkova-Dimitrova, Rusi Rusew, Yuhuan Li, Andrey Popatanasov, Nejc Petek, Martin Štícha and Boris Shivachev
Crystals 2025, 15(9), 772; https://doi.org/10.3390/cryst15090772 - 29 Aug 2025
Viewed by 58
Abstract
Influenza A viruses that cause pandemics, as well as other harmful pathogens (e.g., SARS-CoV-2 variants), are known as the ‘silent bioterrorists’ of the 21st century. Due to high mutability, anti-influenza chemotherapeutic treatment is a vital defense strategy to combat both seasonal and pandemic [...] Read more.
Influenza A viruses that cause pandemics, as well as other harmful pathogens (e.g., SARS-CoV-2 variants), are known as the ‘silent bioterrorists’ of the 21st century. Due to high mutability, anti-influenza chemotherapeutic treatment is a vital defense strategy to combat both seasonal and pandemic influenza strains, especially when vaccines fail. Consequently, the development of novel therapies to combat this serious threat is of great concern. Hence, in this study, 3-(2-thienyl) acrylic acid (TA) was converted into amides of anti-influenza drugs (aminoadamantanes and oseltamivir) through TBTU-mediated coupling. The crystal structures of the thienyl-based amide hybrids (TA-Am (1), TA-Rim (2), TA-Os-OEt (3), and TA-OsC (4)) were also investigated using single-crystal X-ray diffraction, powder X-ray diffraction (PXRD), and differential scanning calorimetry (DSC). Moreover, the antiviral activities of the hybrids against influenza virus A/Fort Monmouth/1/1947 (H1N1), clinically isolated influenza strain A/Wuhan/359/1995 (H3N2), and oseltamivir-resistant A/Jinnan/15/2009 (H1N1) were evaluated in vitro. Amongst the tested thienyl-based amides, bisamide 8 (Boc-Os-Hda-TA) exhibited the most potent activity against influenza virus A (A/Wuhan/359/1995) with an IC50 value of 18.52 μg/mL and a selectivity index (SI) = 13.0. Full article
(This article belongs to the Special Issue Celebrating the 10th Anniversary of International Crystallography)
32 pages, 6749 KB  
Article
Cement Carbonation Under Fermentation Conditions as a Tool for CO2 Emission Management—Technological, Environmental and Economic Analysis
by Michał Pyzalski, Michał Juszczyk, Karol Durczak, Dariusz Sala, Joanna Duda, Marek Dudek and Leonas Ustinovičius
Energies 2025, 18(17), 4588; https://doi.org/10.3390/en18174588 - 29 Aug 2025
Viewed by 89
Abstract
The aim of this study is an interdisciplinary assessment of the potential of cement pastes to permanently bind carbon dioxide (CO2) under anaerobic digestion conditions, considering technological, microstructural, environmental, and economic aspects. The research focused on three types of Portland cement: [...] Read more.
The aim of this study is an interdisciplinary assessment of the potential of cement pastes to permanently bind carbon dioxide (CO2) under anaerobic digestion conditions, considering technological, microstructural, environmental, and economic aspects. The research focused on three types of Portland cement: CEM I 52.5N, CEM I 42.5R-1, and CEM I 42.5R-2, differing in phase composition and reactivity, which were evaluated in terms of their carbonation potential and resistance to chemically aggressive environments. The cement pastes were prepared with a water-to-cement ratio of 0.5 and subjected to 90-day exposure in two environments: a reference environment (tap water) and a fermentation environment (aqueous suspension of poultry manure simulating biogas reactor conditions). XRD, TG/DTA, SEM/EDS, and mercury intrusion porosimetry were applied to analyze CO2 mineralization, phase changes, and microstructural evolution. XRD results revealed a significant increase in calcite content (e.g., for CEM I 52.5N from 5.9% to 41.1%) and the presence of vaterite (19.3%), indicating intense carbonation under organic conditions. TG/DTA analysis confirmed a reduction in portlandite and C-S-H phases, suggesting their transformation into stable carbonate forms. SEM observations and EDS analysis revealed well-developed calcite crystals and the dominance of Ca, C, and O, confirming effective CO2 binding. In control samples, hydration products predominated without signs of mineralization. The highest sequestration potential was observed for CEM I 52.5N, while cements with higher C3A content (e.g., CEM I 42.5R-2) exhibited lower chemical resistance. The results confirm that carbonation under fermentation conditions may serve as an effective tool for CO2 emission management, contributing to improved durability of construction materials and generating measurable economic benefits in the context of climate policy and the EU ETS. The article highlights the need to integrate CO2 sequestration technologies with emission management systems and life cycle assessment (LCA) of biogas infrastructure, supporting the transition toward a low-carbon economy. Full article
(This article belongs to the Special Issue Challenges and Research Trends of Carbon Dioxide Capture)
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19 pages, 3792 KB  
Article
Biological and Genomic Insights into Fusarium acuminatum Causing Needle Blight in Pinus tabuliformis
by Linin Song, Yuying Xu, Tianjin Liu, He Wang, Xinyue Wang, Changxiao Fu, Xiaoling Xie, Yakubu Saddeeq Abubakar, Abah Felix, Ruixian Yang, Xinhong Jing, Guodong Lu, Jiandong Bao and Wenyu Ye
J. Fungi 2025, 11(9), 636; https://doi.org/10.3390/jof11090636 - 29 Aug 2025
Viewed by 206
Abstract
Chinese pine, Pinus tabuliformis, is one of the most important garden plants in northern China, and the planting of this species is of great significance for the improvement of the ecological environment. In this study, different fungi were isolated and purified from [...] Read more.
Chinese pine, Pinus tabuliformis, is one of the most important garden plants in northern China, and the planting of this species is of great significance for the improvement of the ecological environment. In this study, different fungi were isolated and purified from diseased Pinus tabuliformis samples collected in Xi’an city, Shaanxi Province. Of these fungal isolates, only one (isolate AP-3) was pathogenic to the healthy host plant. The pathogenic isolate was identified as Fusarium acuminatum by morphological characteristics and ITS and TEF-1α sequence analyses. The optimal growth conditions for this isolate were further analyzed as follows: Optimal temperature of 25 °C, pH of 11, soluble starch and sodium nitrate as the most preferred carbon and nitrogen sources, respectively. By combining Oxford Nanopore Technologies (ONT) long-read sequencing with Illumina short-read sequencing technologies, we obtained a 41.50 Mb genome assembly for AP-3, with 47.97% GC content and 3.04% repeats. This consisted of 14 contigs with an N50 of 4.64 Mb and a maximum length of 6.45 Mb. The BUSCO completeness of the genome assembly was 98.94% at the fungal level and 97.83% at the Ascomycota level. The genome assembly contained 13,408 protein-coding genes, including 421 carbohydrate-active enzymes (CAZys), 120 cytochrome P450 enzymes (CYPs), 3185 pathogen-host interaction (PHI) genes, and 694 candidate secreted proteins. To our knowledge, this is the first report of F. acuminatum causing needle blight of P. tabuliformis. This study not only uncovered the pathogen responsible for needle blight of P. tabuliformis, but also provided a systematic analysis of its biological characteristics. These findings provide an important theoretical basis for disease control in P. tabuliformis and pave the way for further research into the fungal pathogenicity mechanisms and management strategies. Full article
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25 pages, 2652 KB  
Article
Establishment of Callus Cultures from Dalbergia sissoo Leaf Explants for Production of Skin Therapeutics: An In Vitro and In Silico Study
by Jyotsana and Promila Gupta
Molecules 2025, 30(17), 3531; https://doi.org/10.3390/molecules30173531 - 29 Aug 2025
Viewed by 209
Abstract
Dalbergia sissoo is a commercially exploited timber tree also known for its varied phytochemical constituents holding significant importance in folk medicines with documented biological properties. The present study reports the establishment of callus cultures from its leaf explants for the in vitro production [...] Read more.
Dalbergia sissoo is a commercially exploited timber tree also known for its varied phytochemical constituents holding significant importance in folk medicines with documented biological properties. The present study reports the establishment of callus cultures from its leaf explants for the in vitro production of skin therapeutics. The growth parameters of the callus cultures were calculated. The antioxidant potential of the methanolic extracts of leaf and its callus cultures was evaluated through DPPH assay. Calli at third subculture stage showed the highest antioxidant potential (IC50 273 ± 14.14 µg/mL). A comparative analysis of phytochemical composition was performed using Gas Chromatography–Mass Spectrometry (GC-MS) which revealed the presence of potential skin therapeutic compounds. Out of 146 compounds, only 15 are unique to leaf explants, with the rest being produced in callus cultures. ADME predictions of potential compounds showed their drug likeness properties. The molecular docking of selected phytochemicals such as Chondrillasterol, Stearic acid, and n-Hexadecanoic acid against the tyrosinase enzyme showed better binding affinities than the reference drug (Kojic acid). Molecular dynamics simulation also showed stable conformations of the docked complexes with the target protein. Overall, these investigations unveil for the first time the successful in vitro production of skin therapeutics from D. sissoo, ensuring the sustainable and conservation-friendly utilization of its biomass for medicinal purposes. Full article
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