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12 pages, 3188 KB  
Article
Gene Mapping and Molecular Marker Development for Controlling Purple-Leaf Trait in Pakchoi (Brassica rapa subsp. chinensis (L.) Hanelt)
by Bo Song, Qinyu Yang, Wenqi Zhang, Xiao Yang, Li Zhang, Lin Ouyang, Limei He, Longzheng Chen, Zange Jing, Tao Huang, Hai Xu, Yuejian Li and Qichang Yang
Genes 2025, 16(10), 1184; https://doi.org/10.3390/genes16101184 - 12 Oct 2025
Abstract
Purple pakchoi (Brassica rapa subsp. chinensis (L.) Hanelt) is rich in anthocyanins, which contribute to its significant edible, ornamental, and potential health-promoting value. The previous research on the purple-leaf trait of pakchoi was rather insufficient, key gene controlling the purple-leaf trait remains [...] Read more.
Purple pakchoi (Brassica rapa subsp. chinensis (L.) Hanelt) is rich in anthocyanins, which contribute to its significant edible, ornamental, and potential health-promoting value. The previous research on the purple-leaf trait of pakchoi was rather insufficient, key gene controlling the purple-leaf trait remains to be further elucidated. Fine mapping of the genes responsible for the purple-leaf trait is essential for establishing molecular marker-assisted breeding and facilitating genetic improvement. In this study, we used the inbred purple-leaf line ‘PQC’ and green-leaf line ‘HYYTC’ as parents to construct a six-generation genetic segregation population. We analyzed the inheritance pattern of the purple-leaf trait and combined Bulked Segregant Analysis Sequencing (BSA-seq) with penta-primer amplification refractory mutation system (PARMS) to map the causal gene. The main findings are as follows: the purple-leaf trait is controlled by a single dominant gene. Using BSA-seq and PARMS, the genes were mapped to a 470 kb region (31.18–31.65 Mb) on chromosome A03. Within this interval, 29 candidate genes were identified. Bra017888 which encoding trehalose phosphate synthase 10 (TPS10), was highlighted as a potential regulator of anthocyanin biosynthesis. A developed molecular marker, SNP31304070, based on the final candidate region, successfully distinguished between purple homozygous and purple heterozygous plants in the F2 and F3 populations, showing complete co-segregation with the trait. The marker could be applied to molecular-assisted breeding in purple pakchoi. Full article
(This article belongs to the Section Plant Genetics and Genomics)
22 pages, 3799 KB  
Article
Protein Corona Stability and Removal from PET Microplastics: Analytical and Spectroscopic Evaluation in Simulated Intestinal Conditions
by Tamara Lujic, Tamara Mutic, Ana Simovic, Tamara Vasovic, Stefan Ivanovic, Maja Krstic Ristivojevic, Vesna Jovanovic and Tanja Cirkovic Velickovic
Foods 2025, 14(20), 3454; https://doi.org/10.3390/foods14203454 - 10 Oct 2025
Viewed by 259
Abstract
Microplastics entering the gastrointestinal environment rapidly acquire protein coronas that alter their surface chemistry and analytical detectability. We investigated the physicochemical interactions between fluorescently labeled bovine serum albumin (BSA) and polyethylene terephthalate (PET) microplastics during simulated intestinal exposure and evaluated the stability of [...] Read more.
Microplastics entering the gastrointestinal environment rapidly acquire protein coronas that alter their surface chemistry and analytical detectability. We investigated the physicochemical interactions between fluorescently labeled bovine serum albumin (BSA) and polyethylene terephthalate (PET) microplastics during simulated intestinal exposure and evaluated the stability of the resulting hard corona. Using fluorescence tracking, SDS-PAGE, and FTIR spectroscopy, we showed that BSA forms a persistent corona that resists oxidative-only treatments. Only a combination of oxidation with an alkaline (KOH) or surfactant step (SDS) effectively removed the corona. None of the protocols applied affected polymer integrity. Residual protein in less effective protocols did not show changes on PET spectra in ATR FTIR. To validate the protocol under physiologically relevant complexity, we extended it to PET incubated with single digestive enzymes. FTIR spectra confirmed the removal of protein-specific signals in both systems, with no degradation of PET ester or aromatic functional groups nor signals of protein–polymer interactions. Our results highlight the robustness of protein–PET interactions in biological conditions and provide a variety of protocols for protein corona removal, suitable for diverse applications of microplastic analysis and toxicological studies. Full article
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56 pages, 7355 KB  
Review
Carbon Nanomaterial-Based Electrochemical Biosensors for Alzheimer’s Disease Biomarkers: Progress, Challenges, and Future Perspectives
by Berfin Şak, Helena B. A. Sousa and João A. V. Prior
Biosensors 2025, 15(10), 684; https://doi.org/10.3390/bios15100684 - 9 Oct 2025
Viewed by 421
Abstract
Alzheimer’s disease (AD) requires early and accurate identification of affected brain regions, which can be achieved through the detection of specific biomarkers to enable timely intervention. Carbon nanomaterials (CNMs), including graphene derivatives, carbon nanotubes, graphitic carbon nitride, carbon black, fullerenes, and carbon dots, [...] Read more.
Alzheimer’s disease (AD) requires early and accurate identification of affected brain regions, which can be achieved through the detection of specific biomarkers to enable timely intervention. Carbon nanomaterials (CNMs), including graphene derivatives, carbon nanotubes, graphitic carbon nitride, carbon black, fullerenes, and carbon dots, offer high conductivity, large electroactive surface area, and versatile surface chemistry that enhance biosensor performance. While such properties benefit a wide range of transduction principles (e.g., electrochemical, optical, and plasmonic), this review focuses on their role in electrochemical biosensors. This review summarizes CNM-based electrochemical platforms reported from 2020 to mid-2025, employing aptamers, antibodies, and molecularly imprinted polymers for AD biomarker detection. Covered topics include fabrication strategies, transduction formats, analytical performance in complex matrices, and validation. Reported devices achieve limits of detection from the femtomolar to picogram per milliliter range, with linear ranges typically spanning 2–3 orders of magnitude (e.g., from femtomolar to picomolar, or from picogram to nanogram per milliliter levels). They exhibit high selectivity against common interferents such as BSA, glucose, uric acid, ascorbic acid, dopamine, and non-target peptides, along with growing capabilities for multiplexing and portable operation. Remaining challenges include complex fabrication, limited long-term stability and reproducibility data, scarce clinical cohort testing, and sustainability issues. Opportunities for scalable production and integration into point-of-care workflows are outlined. Full article
(This article belongs to the Special Issue Nano/Micro Biosensors for Biomedical Applications (2nd Edition))
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23 pages, 5760 KB  
Article
Effect of Crosslinking Using Heat on the Physicochemical Features of Bsa–Capsaicin Nanoparticles
by Lino Sánchez-Segura, Silvio Zaina, Angela F. Kú-González, José Alfredo Guzmán-López, Laura E. Zavala-García and Mercedes G. López
Pharmaceutics 2025, 17(10), 1306; https://doi.org/10.3390/pharmaceutics17101306 - 8 Oct 2025
Viewed by 311
Abstract
Background/Objectives: The synthesis of protein nanoparticles (NPs) using the coacervation method is influenced by critical parameters. The use of glutaraldehyde limits the pharmacological applications of NPs in humans due to the potential toxicity of residual aldehydes that remain after the purification of [...] Read more.
Background/Objectives: The synthesis of protein nanoparticles (NPs) using the coacervation method is influenced by critical parameters. The use of glutaraldehyde limits the pharmacological applications of NPs in humans due to the potential toxicity of residual aldehydes that remain after the purification of the nanoparticles. The aim was to assess heat effect as a crosslinking agent for the synthesis of bovine serum albumin (BSA)–capsaicin nanoparticles and its effect on the physicochemical characteristics of nanoparticles. Results: The initial concentrations of BSA and capsaicin in the formulation were directly correlated with the amount of BSA that was transformed into nanoparticles and the loaded capsaicin (r = 0.97, p = 0.0003 and r = 0.95, p = 0.0003), respectively. Furthermore, the morphometric parameters of nanoparticles were affected by the increase in capsaicin concentration, but not by temperature. The nanoparticles increased in dimensions and showed a loss of shape due to coalescence between nanoparticles. The ζ-potential decreased with the increase in the concentration of capsaicin added. This effect compromised the stability of the nanoparticles; on the other hand, molecular interactions were observed between hydrophobic residues of phenylalanine and tyrosine in BSA and the hydrophobic moiety of capsaicin. At the same time, BSA nanoparticles showed a potential for disassembling and delivering the payload capsaicin, which caused an antisteatotic effect in the liver of a murine model. Conclusions: heat (70 °C) can replace crosslinking agents, such as glutaraldehyde. This property is particularly useful when an aldehyde-free synthesis of BSA nanoparticles is needed. Full article
(This article belongs to the Section Drug Delivery and Controlled Release)
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14 pages, 631 KB  
Article
The Impact of Obesity on the Left Atrium and Arrhythmia Recurrence in Patients with Atrial Fibrillation Undergoing Ablation
by Beata Uziębło-Życzkowska, Marek Kiliszek, Krystian Krzyżanowski and Paweł Krzesiński
J. Clin. Med. 2025, 14(19), 7043; https://doi.org/10.3390/jcm14197043 - 5 Oct 2025
Viewed by 328
Abstract
Objectives: Obesity and atrial fibrillation (AF) are strongly linked and are both associated with significant left atrial (LA) pathology. This study aimed to assess differences in LA size and function between obese and non-obese AF patients and to evaluate AF recurrence in [...] Read more.
Objectives: Obesity and atrial fibrillation (AF) are strongly linked and are both associated with significant left atrial (LA) pathology. This study aimed to assess differences in LA size and function between obese and non-obese AF patients and to evaluate AF recurrence in both groups. Materials and Methods: We retrospectively analyzed patients undergoing first-time ablation for AF. Obesity was defined as body mass index ≥30 kg/m2, and patients were divided accordingly into obese and non-obese groups. Results: Among 672 patients (median age of 66 years; 39.1% women), 308 (45.8%) were obese. Obese patients had significantly larger LA dimensions (LA area, LA volume, and LAVI indexed to height2 (but not that indexed to body surface area (BSA)); p < 0.001), as well as higher LA-pressure-related parameters (LA stiffness index (p = 0.004), E-wave velocity (p = 0.002), and E/e′ ratio (p < 0.001)) and invasively measured mean LA pressure (p < 0.0001). However, there were no significant differences in parameters directly reflecting LA function, such as LA emptying fraction, LA reservoir strain, or LA appendage velocity. These findings remained consistent in the sinus rhythm subgroup (n = 374). The 1-year AF recurrence rate did not differ between obese and non-obese groups (data available for 73.8% (496) patients; p = 0.40), regardless of baseline rhythm. Conclusions: In AF patients undergoing their first ablation, obesity was associated with a larger LA size and higher LA pressure. In obese individuals, indexing LA dimensions to height2 seems to better reflect LA enlargement than indexing to BSA. LA function and AF recurrence rates after a 1-year follow-up period were similar between obese and non-obese patients. Full article
(This article belongs to the Special Issue Clinical Aspects of Cardiac Arrhythmias and Arrhythmogenic Disorders)
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14 pages, 3243 KB  
Article
Fine-Mapping of a Red-Skinned Taproot Gene in Radish (Raphanus sativus L.)
by Zhao Liu, Zhenzhen Li, Gaizhen Li and Linyi Qiao
Plants 2025, 14(19), 3065; https://doi.org/10.3390/plants14193065 - 3 Oct 2025
Viewed by 345
Abstract
The skin color of radish taproots is an important commodity character that directly affects the choice behavior of consumers. Here, we identified a skin color gene carried by a red-skinned inbred line, SXAU-R2. Genetic population was constructed by the crossing of SXAU-R2 and [...] Read more.
The skin color of radish taproots is an important commodity character that directly affects the choice behavior of consumers. Here, we identified a skin color gene carried by a red-skinned inbred line, SXAU-R2. Genetic population was constructed by the crossing of SXAU-R2 and a white-skinned inbred line, SXAU-W2, and the taproots of F1 plants exhibited intermediate color. In the F2 population, the separation ratio of taproot skin color indicated that the phenotype was controlled by one major locus, named RST1 (Red-Skinned Taproot 1). Combined with bulked segregant analysis and RNA sequencing (BSA-seq), 2640 single nucleotide polymorphisms (SNPs) were detected between the annotated genes of the red skin bulk and white skin bulk. Molecular markers were developed in the SNP-enriched 27~32 Mbp region of chromosome 7, and then RST1 was mapped in the genetic interval between flanking markers SSR-14 and SSR-22. Using F2:3 lines derived from a key F2 heterozygote, RST1 was narrowed down into a 530 Kbp interval. There were 46 expressed annotated genes in the fine-mapping region, and a gene encoding MYB was selected as the candidate of RST1. Finally, based on Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analysis and RT-qPCR, we identified the potential interacting genes RsbHLH and RsWD, as well as the latent target genes RsDFR and RsANS of RST1 in the anthocyanin synthesis pathway. These results provide an understanding of the genetic mechanisms regulating anthocyanin synthesis and offer an efficient molecular marker for the radish breeding of skin color. Full article
(This article belongs to the Special Issue Genetic Mapping of Agronomic Traits in Crops)
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20 pages, 3589 KB  
Article
Comparison of Different Aliphatic Polyester-Based Microparticles as Protein Delivery Systems
by Viktor Korzhikov-Vlakh, Ekaterina Sinitsyna, Mariia Stepanova, Evgenia Korzhikova-Vlakh and Tatiana Tennikova
Polymers 2025, 17(19), 2676; https://doi.org/10.3390/polym17192676 - 3 Oct 2025
Viewed by 301
Abstract
The utilization of encapsulated biopharmaceuticals, including peptides and proteins, has grown substantially in recent years. In this study, the influence of aliphatic polyester physicochemical properties, specifically crystallinity and hydrophobicity, on the development of protein-loaded microparticles was investigated. A series of polyesters, namely amorphous [...] Read more.
The utilization of encapsulated biopharmaceuticals, including peptides and proteins, has grown substantially in recent years. In this study, the influence of aliphatic polyester physicochemical properties, specifically crystallinity and hydrophobicity, on the development of protein-loaded microparticles was investigated. A series of polyesters, namely amorphous PDLLA and semicrystalline PLLA, PCL, and PPDL, were synthesized via chemical and enzymatic ring-opening polymerization. Bovine serum albumin (BSA)-loaded microparticles were fabricated using a water-in-oil-in-water (w/o/w) double emulsion solvent evaporation method. The size of microparticles obtained was determined by scanning electron microscopy and dynamic light scattering methods. The enzymatic degradation of the polymer microparticles was assessed through incubation in a lipase-containing buffer solution. BSA and α-chymotrypsin (ACHT) were used as model proteins for the preparation of encapsulated polymer microspheres and comparison of their characteristics and properties. Protein encapsulation efficacy, release rate, and enzyme activity retained after encapsulation were evaluated and compared for selected aliphatic polyesters. The release profiles were processed with the use of various mathematical models to reveal the possible mechanism(s) of protein release. Full article
(This article belongs to the Special Issue Polyester-Based Materials: 3rd Edition)
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20 pages, 3311 KB  
Article
Research and Development of pH-Sensitive Delivery Systems for Protein Molecule Delivery Based on Chitosan and Hydroxyapatite
by Elina E. Chernikova, Maxim D. Zagvozkin, Aleksander A. Buzaev, Irina A. Kurzina and Svetlana A. Ulasevitch
J. Compos. Sci. 2025, 9(10), 525; https://doi.org/10.3390/jcs9100525 - 1 Oct 2025
Viewed by 259
Abstract
The degree of bovine serum albumin (BSA) released from particles based on chitosan and hydroxyapatite prepared by encapsulation and chemisorption methods, crosslinked by aldol condensation, was studied. The obtained materials’ composition was qualitatively determined by IR spectroscopy; phase identification and surface morphology were [...] Read more.
The degree of bovine serum albumin (BSA) released from particles based on chitosan and hydroxyapatite prepared by encapsulation and chemisorption methods, crosslinked by aldol condensation, was studied. The obtained materials’ composition was qualitatively determined by IR spectroscopy; phase identification and surface morphology were analyzed by X-ray diffraction and scanning electron microscopy, respectively. A spectrophotometric method was used to quantitatively assess the loading and release degree of encapsulated/chemisorbed BSA from polymer microspheres. To determine the release mechanism, the data on the amount of BSA released were analyzed according to the zero- and first-order, Higuchi, and Korsmayer–Peppas models. Full article
(This article belongs to the Section Biocomposites)
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13 pages, 2180 KB  
Article
Mutation in the LONGIFOLIA1 Gene Resulted in Suppressed Insensitivity of Arabidopsis thaliana proteolysis6 Mutant to Ethylene During Seed Germination
by Xu Wang, Ying Luo, Yuan Cao, Yujin Gong, Francoise Corbineau and Yong Xiang
Seeds 2025, 4(4), 48; https://doi.org/10.3390/seeds4040048 - 30 Sep 2025
Viewed by 272
Abstract
Seed dormancy and germination is regulated by internal hormones and exogenous environment cues. Ethylene is one of the hormones that break seed dormancy and induce seed germination. Our previous study showed that N-degron pathway gene, proteolysis6 (PRT6) was involved in dormancy [...] Read more.
Seed dormancy and germination is regulated by internal hormones and exogenous environment cues. Ethylene is one of the hormones that break seed dormancy and induce seed germination. Our previous study showed that N-degron pathway gene, proteolysis6 (PRT6) was involved in dormancy release by ethylene, the defection of which exhibiting ethylene-insensitivity in Arabidopsis thaliana. In the present study, through screening an ethyl methyl sulfonate-mutagenized (EMS) population of prt61, we isolated a recessive mutant that acted as a suppressor of prt6 that rescued its insensitivity to ethylene as well as a phenotype of shorter silique length. Further bulk segregant analysis on F2 population identified a premature termination located in the third exon of LONGIFOLIA1 (LNG1), previously reported in the regulation of longitudinal cell elongation. Mutation of LNG1 in prt61 background by CRISPR-Cas9 confirmed that LNG1 was epistatic to PRT6 in seed responsiveness to ethylene. Our finding proposed the pleiotropic effect of LNG1 in seed dormancy breakage by ethylene via PRT6, providing novel functional component at the downstream of the coordinated PRT6 and ethylene signaling pathway. Full article
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24 pages, 616 KB  
Article
Pre-Treatment Nutritional Status as a Predictor of Clinical Outcomes in Moderate-to-Severe Plaque Psoriasis Patients Undergoing Cyclosporine A Therapy
by Wojciech Kulej, Beniamin Oskar Grabarek, Martyna Stefaniak, Laura Opalska, Piotr Michalski, Aleksandra Plata-Babula and Anna Michalska-Bańkowska
Nutrients 2025, 17(19), 3098; https://doi.org/10.3390/nu17193098 - 29 Sep 2025
Viewed by 230
Abstract
Background/Objectives: Psoriasis is a chronic immune-mediated disease frequently accompanied by systemic inflammation and metabolic disturbances. Nutrition plays a crucial role in modulating inflammatory pathways, yet the impact of baseline dietary status on systemic therapy outcomes remains underexplored. Methods: A total of [...] Read more.
Background/Objectives: Psoriasis is a chronic immune-mediated disease frequently accompanied by systemic inflammation and metabolic disturbances. Nutrition plays a crucial role in modulating inflammatory pathways, yet the impact of baseline dietary status on systemic therapy outcomes remains underexplored. Methods: A total of 37 patients (20 men, 17 women; mean age 47.8 ± 4.87 years) scheduled for cyclosporine A (CsA) therapy underwent dietary assessment using 24 h recall and food frequency questionnaires. Intake was compared with dietary reference values. Psoriasis severity was measured by using the Psoriasis Area and Severity Index (PASI) and Body Surface Area (BSA) at baseline, day 42, and day 84. Mixed-effects regression models adjusted for body mass index (BMI), age, and sex assessed associations between nutrient adequacy and clinical outcomes. Results: Participants exhibited frequent dietary imbalances, including low polyunsaturated fatty acids, fiber, vitamin D, folate, and minerals such as magnesium and zinc, alongside excess saturated fat and sodium. Adequate intake of fiber, eicosapentaenoic acid (EPA)+ docosahexaenoic acid (DHA), and vitamins A and D, folate, magnesium, and zinc was independently associated with a lower baseline PASI/BSA and faster improvement during CsA therapy (p < 0.05). Higher BMI, older age, and male sex predicted poorer outcomes. Conclusions: Pre-treatment nutritional inadequacies are common in psoriasis and independently predict diminished therapeutic response to CsA. Early nutritional optimization may enhance treatment efficacy and support long-term disease control. Integrating dietary assessment in psoriasis management represents a feasible, impactful adjunct to pharmacotherapy. Full article
(This article belongs to the Section Clinical Nutrition)
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23 pages, 4574 KB  
Article
A Heterobimetallic Au(I)–Ru(II) Complex Bridged by dppb: Synthesis, Structural and Solution Characterization, BSA Interaction and In Vivo Toxicity Evaluation in Wistar Rats
by Adnan Zahirović, Sunčica Roca, Muhamed Fočak, Selma Fetahović, Višnja Muzika, Damir Suljević, Anela Topčagić, Maja Mitrašinović-Brulić, Irnesa Osmanković, Debbie C. Crans and Aleksandar Višnjevac
Inorganics 2025, 13(10), 323; https://doi.org/10.3390/inorganics13100323 - 29 Sep 2025
Viewed by 305
Abstract
A novel heterobimetallic ruthenium(II)–gold(I) complex featuring a bridging bis(diphenylphosphino)butane (dppb) ligand was prepared and fully characterized. Single-crystal X-ray diffraction revealed a piano-stool geometry around Ru(II) with η6-cymene, two chlorido ligands, and one phosphorus atom from dppb, while the Au(I) center adopts [...] Read more.
A novel heterobimetallic ruthenium(II)–gold(I) complex featuring a bridging bis(diphenylphosphino)butane (dppb) ligand was prepared and fully characterized. Single-crystal X-ray diffraction revealed a piano-stool geometry around Ru(II) with η6-cymene, two chlorido ligands, and one phosphorus atom from dppb, while the Au(I) center adopts a linear P–Au–Cl coordination. Structural integrity in the solution was confirmed by 1D and 2D NMR spectroscopy, while solution behavior was further monitored by variable solvent 31P NMR and UV/Vis spectroscopy, indicating that the organometallic Ru–arene core remains intact, whereas the chlorido ligands coordinated to Ru exhibit partial lability. Complementary characterization included elemental analysis, FTIR, and UV/Vis spectroscopy. Spectrofluorimetric and FRET analyses showed that Au(dppb), Ru(dppb), and the heterobimetallic AuRu complex bind to BSA with apparent constants of 1.41 × 105, 5.12 × 102, and 2.66 × 104 M−1, respectively, following a static quenching mechanism. In vivo biological evaluation in Wistar rats revealed no significant hepatotoxicity or nephrotoxicity, with only mild and reversible histological alterations and preserved hepatocyte nuclear morphology. Hematological analysis indicated a statistically significant reduction in leukocyte populations, suggesting immunomodulatory potential, while elevated serum glucose levels point to possible endocrine or metabolic activity. These findings highlight compound structural stability and intriguing bioactivity profile, making it a promising platform for further organometallic drug development and testing. Full article
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20 pages, 3628 KB  
Article
A Stable Delivery System for Meretrix meretrix Derived Immunomodulatory Peptide (QLNWD): Fabrication and Characterization of Glycosylated Protein Nanoparticle
by Wanyi Wu, Zhixuan Wu, Jiamin Cai, Wenhong Cao, Haisheng Lin, Jialong Gao, Xiuping Fan, Huina Zheng and Xiaoming Qin
Mar. Drugs 2025, 23(10), 385; https://doi.org/10.3390/md23100385 - 27 Sep 2025
Viewed by 378
Abstract
In this study, nanoparticles prepared by the heat-induced self-assembly of bovine serum albumin-dextran conjugates (BSA-DX) were utilized as an effective delivery system for the immunoregulatory peptide Gln-Leu-Asn-Trp-Asp (QLNWD) from Meretrix meretrix. The effects of dry-heating duration on the fabrication and characteristics of [...] Read more.
In this study, nanoparticles prepared by the heat-induced self-assembly of bovine serum albumin-dextran conjugates (BSA-DX) were utilized as an effective delivery system for the immunoregulatory peptide Gln-Leu-Asn-Trp-Asp (QLNWD) from Meretrix meretrix. The effects of dry-heating duration on the fabrication and characteristics of glycoprotein nanoparticles (GBA NPs) were investigated. Stable GBA NPs (110.84 nm) were obtained after 9 h of dry-heating. Depending on the addition sequence of QLNWD, the QLNWD-loaded nanoparticles were categorized into two types: pre-loading and post-loading. The two strategies were evaluated based on physicochemical characterization, colloidal stability, and RAW264.7 macrophage uptake. Results showed that upon QLNWD incorporation, both pre-loading NPs and post-loading NPs exhibited spherical morphology, with particle sizes decreasing to 105.51 nm and 94.27 nm, respectively. The encapsulation efficiency of pre-loading NPs for QLNWD was higher (87.74%), and the co-localization ability between post-loading NPs and QLNWD was stronger (Pearson’s correlation coefficient = 0.95). In vitro simulated gastrointestinal digestion experiments showed that QLNWD bioaccessibility increased to 47.5% and 42.7% for pre-loaded and post-loaded NPs, respectively. Compared to free QLNWD, NP encapsulation significantly enhanced the uptake of QLNWD by macrophages. Thus, GBA NPs, particularly those prepared by the pre-loading method, are considered promising delivery systems for marine bioactive peptides. Full article
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13 pages, 621 KB  
Article
5-Hydroxymethylfurfural: A Particularly Harmful Molecule Inducing Toxic Lipids and Proteins?
by Joachim Greilberger, Georg Feigl, Matthias Greilberger, Simona Bystrianska and Michaela Greilberger
Molecules 2025, 30(19), 3897; https://doi.org/10.3390/molecules30193897 - 26 Sep 2025
Viewed by 313
Abstract
Introduction: 5-HMF is a molecule found in carbohydrate-rich foods that is associated not only with cancer and anaphylactic reactions, but also with anti-oxidant properties. Questions arose as to whether 5-HMF exhibited a catalytic effect in relation to lipid peroxidation and lipoprotein oxidation in [...] Read more.
Introduction: 5-HMF is a molecule found in carbohydrate-rich foods that is associated not only with cancer and anaphylactic reactions, but also with anti-oxidant properties. Questions arose as to whether 5-HMF exhibited a catalytic effect in relation to lipid peroxidation and lipoprotein oxidation in presence of metals and/or radicals. Methods: Peroxynitrite (ONOO)-induced chemiluminescence and ONOO nitration of tyrosine residues on BSA using anti-nitro-tyrosine-antibodies were used to measure the protection of 5-HMF against peroxides or nitration compared to vitamin C (VitC). The reductive potential of 5-HMF or VitC on Cu2+ or Fe3 was estimated using the bicinchoninic acid (BCA) or Fenton-complex method. Human plasma was used to measure the generation of malondialdehyde (MDA), 4-hydroxynonenal (HNE), and total thiols after Fe2+/H2O2 oxidation in the presence of different concentrations of 5-HMF or VitC. Finally, Cu2+ oxidation of LDL after 4 h was carried out with 5-HMF or VitC, measuring the concentration of MDA in LDL with the thiobarbituric assay (TBARS). Results: VitC was 4-fold more effective than 5-HMF in scavenging ONOO to nearly 91.5% at 4 mM, with the exception of 0.16 mM, where the reduction of ONOO by VitC was 3.3-fold weaker compared to 0.16 mM 5-HMF. VitC or 5-HMF at a concentration of 6 mM inhibited the nitration of tyrosine residues on BSA to nearly 90% with a similar course. While 5-HMF reduced free Fe3+ in presence of phenanthroline, forming Fe2+ (phenantroleine)3 [Fe2+(phe)3] or complexed Cu2+(BCA)4 to Cu+(BCA)4 weakly, VitC was 7- to 19-fold effective in doing so over all the used concentrations (0–25 mM). A Fe2+—H2O2 solution mixed with human plasma showed a 6–10 times higher optical density (OD) of MDA or HNE in the presence of 5-HMF compared to VitC. The level of thiols was significantly decreased in the presence of higher VitC levels (1 mM: 198.4 ± 7.7 µM; 2 mM: 160.0 ± 13.4 µM) compared to equal 5-HMF amounts (2562 ± 7.8 µM or 242.4 ± 2.5 µM), whereas the usage of lower levels at 0.25 µM 5-HMF resulted in a significant decrease in thiols (272.4 ± 4.0 µM) compared to VitC (312.3 ± 19.7 µM). Both VitC and 5-HMF accelerated copper-mediated oxidation of LDL equally: while the TBARS levels from 4 h oxidized LDL reached 137.7 ± 12.3 nmol/mg, it was 1.7-fold higher using 6 mM VitC (259.9 ± 10.4 nmol/mg) or 6 mM 5-HMF (239.3 ± 10.2 nmol/mg). Conclusions: 5-HMF appeared to have more pro-oxidative potential compared to VitC by causing lipid peroxidation as well as protein oxidation. Full article
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16 pages, 1195 KB  
Article
Insight into the Roles of Albumin—Alone and in Combination with Either Voriconazole or Antimicrobial Peptides Derived from Chromogranin A—In the Growth of Different Microbial Species
by Francis Schneider, Sophie Hellé, Jean-Marc Strub, François-Xavier von Hunolstein, Pierre Schaaf, Philippe Lavalle, Francesco Scavello and Marie-Hélène Metz-Boutigue
Antibiotics 2025, 14(10), 974; https://doi.org/10.3390/antibiotics14100974 - 26 Sep 2025
Viewed by 272
Abstract
Background: Whether therapeutic albumin (ThHSA) can serve as a defense tool in Candida species (spp.) infections is still a matter of debate, although many physicians are in the habit of infusing ThHSA to restore the physiological concentration of endogenous human serum albumin (HSA). [...] Read more.
Background: Whether therapeutic albumin (ThHSA) can serve as a defense tool in Candida species (spp.) infections is still a matter of debate, although many physicians are in the habit of infusing ThHSA to restore the physiological concentration of endogenous human serum albumin (HSA). Given the need for innovative anti-Candida strategies, we assessed in vitro the role of ThHSA alone or in combination with voriconazole (VCZ) in combating Candida spp. growth and the role of bovine serum albumin (BSA)—used as a substitute for HSA—with two endogenous bovine antimicrobial peptides in combating C. albicans and other microbes. Results: The combination of ThHSA with VCZ enhanced the antifungal effect on C. albicans, sensitive C. tropicalis, sensitive C. glabrata, and C. lusitaniae. However, for resistant C. tropicalis, the combination of ThHSA with VCZ promoted yeast growth, and VCZ tended to suppress the antimicrobial effect of ThHSA on resistant C. glabrata. As to the possible transposition of ThHSA-type properties to BSA (as regards the growth inhibition of other pathogens), we tested combinations of BSA with two physiological chromogranin A-derived antimicrobial peptides (catestatin and cateslytin). BSA enhanced significantly the activity of catestatin (but not cateslytin) in combating C. albicans, A. fumigatus, and M. luteus, but was inactive against S. aureus and E. coli. Conclusions: Our experiments support the fact that albumins display intrinsic antimicrobial properties, with an unpredictable growth inhibitory effect on various microbes. ThHSA can thus be an adjunctive tool for more efficient care of some, though not all, infections. The interaction of BSA with catestatin and cateslytin is related to their structure, with BSA significantly enhancing the effect of catestatin but not that of cateslytin. Full article
(This article belongs to the Special Issue Bioactive Peptides and Their Antibiotic Activity)
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Article
Development and Characterization of Chitosan Microparticles via Ionic Gelation for Drug Delivery
by Zahra Rajabimashhadi, Annalia Masi, Sonia Bagheri, Claudio Mele, Gianpiero Colangelo, Federica Paladini and Mauro Pollini
Polymers 2025, 17(19), 2603; https://doi.org/10.3390/polym17192603 - 26 Sep 2025
Viewed by 463
Abstract
This study explores the formulation of chitosan microparticles through ionic gelation and presents detailed physicochemical characterization, release studies, and the utility and potential uses for drug delivery. Three formulations were prepared under rate-controlled conditions (stirring at 800 rpm and pH maintained at 4.6) [...] Read more.
This study explores the formulation of chitosan microparticles through ionic gelation and presents detailed physicochemical characterization, release studies, and the utility and potential uses for drug delivery. Three formulations were prepared under rate-controlled conditions (stirring at 800 rpm and pH maintained at 4.6) with and without stabilizers to examine the effects of formulation parameters on particle morphology and structural stability. To determine different structural and chemical characteristics, Attenuated Total Reflectance Fourier-Transform Infrared spectroscopy (ATR–FTIR), Scanning Electron Microscopy (SEM), and dynamic light scattering (DLS) were utilized, which confirmed that the particles formed and assessed size distribution and structural integrity. Atomic force microscopy (AFM) was used to quantify surface roughness and potential nanomechanical differences that may derive from the use of different modifiers. Coformulation of bovine serum albumin (BSA) permitted assessment of encapsulation efficiency and drug release capacity. Based on in vitro release evidence, the protein released at a different rate, and the dispersion of formulations under physiological conditions (PBS, pH 7.4, 37 °C) confirmed the differences in stability between formulations. The tunable physical characteristics, mild fabrication conditions, and controlled drug release demonstrated that the chitosan particles could have useful relevance as a substrate for localized drug delivery and as a bioactive scaffold for tissue regenerative purposes. Full article
(This article belongs to the Special Issue Advanced Polymeric Biomaterials for Drug Delivery Applications)
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