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11 pages, 2386 KiB  
Article
Genome-Wide In Silico Analysis Expanding the Potential Allergen Repertoire of Mango (Mangifera indica L.)
by Amit Singh, Aayan Zarif, Annelise N Huynh, Zhibo Yang and Nagib Ahsan
Appl. Sci. 2025, 15(15), 8375; https://doi.org/10.3390/app15158375 - 28 Jul 2025
Viewed by 242
Abstract
The potential of a protein to cause an allergic reaction is often assessed using a variety of computational techniques. Leveraging advances in high-throughput protein sequence data coupled with in silico or computational methods can be used to systematically analyze large proteomes for allergenic [...] Read more.
The potential of a protein to cause an allergic reaction is often assessed using a variety of computational techniques. Leveraging advances in high-throughput protein sequence data coupled with in silico or computational methods can be used to systematically analyze large proteomes for allergenic potential. Despite mango’s widespread consumption and growing clinical reports of hypersensitivity, the full extent of their allergenicity is yet unknown. In this study, for the first time, we conducted a genome-wide in silico analysis by analyzing a total of 54,010 protein sequences to identify the complete spectrum of potential mango allergens. These proteins were analyzed using various bioinformatics tools to predict their allergenic potential based on sequence similarity, structural features, and known allergen databases. In addition to the known mango allergens, including Man i 1, Man i 2, and Man i 3, our findings demonstrated that several isoforms of cysteine protease, non-specific lipid-transfer protein (LTP), legumin B-like, 11S globulin, vicilin, thaumatin-like protein, and ervatamin-B family proteins exhibited strong allergenic potential, with >80% 3D epitope identity, >70% linear 80 aa window identity, and matching with >80 known allergens. Thus, a genome-wide in silico study provided a comprehensive profile of the possible mango allergome, which could help identify the low-allergen-containing mango cultivars and aid in the development of accurate assays for variety-specific allergic reactions. Full article
(This article belongs to the Special Issue New Diagnostic and Therapeutic Approaches in Food Allergy)
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23 pages, 3587 KiB  
Article
Anti-Trypanosoma cruzi Potential of New Pyrazole-Imidazoline Derivatives
by Edinaldo Castro de Oliveira, Leonardo da Silva Lara, Lorraine Martins Rocha Orlando, Sarah da Costa Lanera, Thamyris Perez de Souza, Nathalia da Silva Figueiredo, Vitoria Barbosa Paes, Ana Carolina Mazzochi, Pedro Henrique Myra Fernandes, Maurício Silva dos Santos and Mirian Claudia de Souza Pereira
Molecules 2025, 30(15), 3082; https://doi.org/10.3390/molecules30153082 - 23 Jul 2025
Viewed by 381
Abstract
Chagas disease, caused by Trypanosoma cruzi, poses a significant public health challenge due to its widespread prevalence, limited therapeutic options, and adverse effects associated with available medications. In this study, we developed 13 novel pyrazole-imidazoline derivatives, inspired by a previously identified cysteine [...] Read more.
Chagas disease, caused by Trypanosoma cruzi, poses a significant public health challenge due to its widespread prevalence, limited therapeutic options, and adverse effects associated with available medications. In this study, we developed 13 novel pyrazole-imidazoline derivatives, inspired by a previously identified cysteine protease inhibitor, and evaluated their antiparasitic activity. Our in silico analyses predicted favorable physicochemical profiles and promising oral bioavailability for these derivatives. Upon phenotypic screening, we observed that these new derivatives exhibited low cytotoxicity (CC50 > 100 µM) and marked efficacy against intracellular amastigotes. Derivative 1k showed high activity (IC50 = 3.3 ± 0.2 µM), selectivity (SI = 73.9), and potency (pIC50 = 5.4). In a 3D cardiac microtissue model, 1k significantly reduced parasite load, matching the efficacy of benznidazole (Bz) even at lower concentrations. Both 1k and Bz effectively prevented parasite recrudescence; however, neither resulted in parasite sterility under the experimental conditions employed. The combination of 1k–Bz yielded an additive interaction, highlighting its potential for in vivo combination therapy. While structural changes abolished cysteine protease inhibition, incorporating a CF3 substituent at the para position and excluding the amino group enhanced antiparasitic activity. These findings reinforce the promise of the pyrazole-imidazoline scaffold and support further structural optimizations to develop innovative candidates for treating Chagas disease. Full article
(This article belongs to the Special Issue Heterocyclic Compounds for Drug Design and Drug Discovery)
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16 pages, 2096 KiB  
Article
Environmental Antidepressants Disrupt Metabolic Pathways in Spirostomum ambiguum and Daphnia magna: Insights from LC-MS-Based Metabolomics
by Artur Jędreas, Sylwia Michorowska, Agata Drobniewska and Joanna Giebułtowicz
Molecules 2025, 30(14), 2952; https://doi.org/10.3390/molecules30142952 - 13 Jul 2025
Viewed by 456
Abstract
Pharmaceuticals such as fluoxetine, paroxetine, sertraline, and mianserin occur in aquatic environments at low yet persistent concentrations due to their incomplete removal in wastewater treatment plants. Although frequently detected, these neuroactive compounds remain underrepresented in ecotoxicological assessments. Given their pharmacodynamic potency, environmentally relevant [...] Read more.
Pharmaceuticals such as fluoxetine, paroxetine, sertraline, and mianserin occur in aquatic environments at low yet persistent concentrations due to their incomplete removal in wastewater treatment plants. Although frequently detected, these neuroactive compounds remain underrepresented in ecotoxicological assessments. Given their pharmacodynamic potency, environmentally relevant concentrations may induce sublethal effects in non-target organisms. In this study, we applied untargeted LC-MS-based metabolomics to investigate the sublethal effects of four widely used antidepressants—paroxetine, sertraline, fluoxetine (SSRIs), and mianserin (TeCA)—on two ecologically relevant freshwater invertebrates: S. ambiguum and D. magna. Organisms were individually exposed to each compound for 48 h at a concentration of 100 µg/L and 25 µg/L, respectively. Untargeted metabolomics captured the sublethal biochemical effects of these antidepressants, revealing both shared disruptions—e.g., in glycerophospholipid metabolism and cysteine and methionine metabolism—and species-specific responses. More pronounced pathway changes observed in D. magna suggest interspecies differences in metabolic capacity or xenobiotic processing mechanisms between taxa. Among the four antidepressants tested, sertraline in D. magna and fluoxetine in S. ambiguum exerted the most extensive metabolomic perturbations, as evidenced by the highest number and pathway impact scores. In D. magna, fluoxetine and mianserin produced similar metabolic profiles, largely overlapping with those of sertraline, whereas paroxetine affected only a single pathway, indicating minimal impact. In S. ambiguum, paroxetine and mianserin elicited comparable responses, also overlapping with those of fluoxetine, while sertraline triggered the fewest changes. These results suggest both compound-specific effects and a conserved metabolic response pattern among the antidepressants used. They also underscore the considerable potential of metabolomics as a powerful and sensitive tool for ecotoxicological risk assessments, particularly when applied across multiple model organisms to capture interspecies variations. However, further research is essential to identify which specific pathway disruptions are most predictive of adverse effects on organismal health. Full article
(This article belongs to the Special Issue Advances in the Mass Spectrometry of Chemical and Biological Samples)
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21 pages, 6590 KiB  
Article
Comparative Analysis of the Complete Chloroplast Genomes of Eight Salvia Medicinal Species: Insights into the Deep Phylogeny of Salvia in East Asia
by Yan Du, Yang Luo, Yuanyuan Wang, Jiaxin Li, Chunlei Xiang and Meiqing Yang
Curr. Issues Mol. Biol. 2025, 47(7), 493; https://doi.org/10.3390/cimb47070493 - 27 Jun 2025
Viewed by 373
Abstract
Salvia, a medicinally and economically important genus, is widely used in traditional medicine, agriculture, and horticulture. This study compares the chloroplast genomes of eight East Asian Salvia species to assess genetic diversity, structural features, and evolutionary relationships. Complete chloroplast genomes were sequenced, [...] Read more.
Salvia, a medicinally and economically important genus, is widely used in traditional medicine, agriculture, and horticulture. This study compares the chloroplast genomes of eight East Asian Salvia species to assess genetic diversity, structural features, and evolutionary relationships. Complete chloroplast genomes were sequenced, annotated, and analyzed for gene content, codon usage, and repetitive sequences. Phylogenetic relationships were reconstructed using Maximum Likelihood, Maximum Parsimony and Bayesian inference. The genomes exhibited a conserved quadripartite structure (151,081–152,678 bp, GC content 37.9–38.1%), containing 114 unique genes with consistent arrangement. Codon usage favored A/T endings, with leucine (Leu) most frequent and cysteine (Cys) least. We identified 281 long sequence repeats (LSRs) and 345 simple sequence repeats (SSRs), mostly in non-coding regions. Comparative analysis revealed five hypervariable regions (trnH-psbA, rbcL-accD, petA-psbJ, rpl32-trnL, ycf1) as potential molecular markers. Phylogenetic analysis confirmed the monophyly of East Asian Salvia, dividing them into five clades, with Sect. Sonchifoliae basal. While G1, G3, and G8 were monophyletic, G5 and G6 were paraphyletic, and the G7-G8 relationship challenged traditional classifications. The genomic evidence provides crucial insights for resolving long-standing taxonomic uncertainties and refining the classification system of Salvia. These findings suggest a complex evolutionary history involving hybridization and incomplete lineage sorting, providing valuable genomic insights for Salvia phylogeny, taxonomy, and conservation. Full article
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21 pages, 6129 KiB  
Article
Diversity and Novelty of Venom Peptides in Vermivorous Cone Snails, Subgenus Rhizoconus (Gastropoda: Mollusca)
by Christine Marie C. Florece, Quentin Kaas, Neda Barghi and Arturo O. Lluisma
Mar. Drugs 2025, 23(7), 266; https://doi.org/10.3390/md23070266 - 26 Jun 2025
Viewed by 611
Abstract
A large majority of cone snails (a species in the genus Conus) are vermivorous (worm-hunting), but the diversity and bioactivity of their venom peptides remain largely unexplored. In this study, we report the first venom gland transcriptomes from two species in the [...] Read more.
A large majority of cone snails (a species in the genus Conus) are vermivorous (worm-hunting), but the diversity and bioactivity of their venom peptides remain largely unexplored. In this study, we report the first venom gland transcriptomes from two species in the Rhizoconus clade, Conus capitaneus and Conus mustelinus, and a new Conus miles transcriptome from a specimen collected in the Philippines. From the set of assembled sequences, a total of 225 C. capitaneus, 121 C. miles, and 168 C. mustelinus putative peptide toxin transcripts were identified, which were assigned to 27 canonical gene superfamilies in C. capitaneus and 24 in C. miles and in C. mustelinus. Most of these venom peptides are novel, and some exhibit new cysteine patterns. Clustering also revealed 12 putative novel gene superfamilies, highlighting the diversity of uncharacterized venom peptides in this group. The O1-, M-, O2-, and con-ikot-ikot superfamilies were the most abundant, while gene superfamilies such as D and G2 were highly expressed. Several hormone-like conopeptides were also identified in this study, revealing the vast diversity of conopeptides from the Rhizoconus species. Full article
(This article belongs to the Section Marine Toxins)
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13 pages, 703 KiB  
Article
Proteolytic Activity Inhibition in Gingival Fluid by Cysteine Protease Inhibitors Obtained from Egg White and Fallopia japonica Extract: An In Vitro Study
by Maciej Siewiński, Maciej Dobrzyński, Krzysztof Gołąb, Maciej Janeczek, Łukasz Nieradko, Barbara Bażanów, Andrzej Rapak, Marius Boariu, Stefan-Ioan Stratul, Alla Belova, Sorina Mihaela Solomon, Renata Samulak and Monika Machoy
Biomedicines 2025, 13(7), 1545; https://doi.org/10.3390/biomedicines13071545 - 25 Jun 2025
Viewed by 388
Abstract
Background/Objectives: Gingipains produced by P. gingivalis have been shown to be directly related to periodontal tissue degradation and are significant molecular targets in therapy of periodontitis. Blocking the activity of these enzymes should reduce survival of this pathogen and mitigate the effects [...] Read more.
Background/Objectives: Gingipains produced by P. gingivalis have been shown to be directly related to periodontal tissue degradation and are significant molecular targets in therapy of periodontitis. Blocking the activity of these enzymes should reduce survival of this pathogen and mitigate the effects of inflammation in periodontitis. Therefore, gingipains inhibitors and specific antibodies could be recommended in the treatment of periodontitis. Cysteine peptidase inhibitors can be obtained by chemical synthesis, or isolated from natural raw materials. This research has the following aims: 1. to analyze in vitro the inhibition of cysteine protease activity in the gingival crevicular fluid (GCF) and 2. to compare the toxicity of natural raw inhibitors (obtained from Fallopia japonica plant and egg white) with chlorhexidine (CHX) using an MTS viability test. Methods: Samples of GCF were collected from healthy (N = 17) individuals and (N = 65) periodontal patients. Cysteine peptidase activity was inhibited by adding a solution of cystatin from egg white (with 20% glycerol), or cystatin from knotweed, or low molecular weight inhibitors (MW < 3 kDa) from egg white and knotweed against Nα-Benzoyl-DL-arginine 4-nitroanilide hydrochloride. Results: There was a statistically significant difference between the inhibition means of cysteine protease activity for the five groups (p < 0.001). Means for the four groups of patients with periodontitis were not statistically significant different from each other (p = 0.320). The inhibition rates were higher in periodontitis patients. The toxicity of knotweed cystatin inhibitor was several times lower than the toxicity of E-64d, and of CHX. Conclusion: Cysteine protease inhibitors isolated from egg or plants were non-toxic, effectively inhibited the activity of cysteine proteases in GCF, and may be a promising alternative to more toxic standard antimicrobials (CHX) in preventing periodontal tissue breakdown. Full article
(This article belongs to the Section Biomedical Engineering and Materials)
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12 pages, 305 KiB  
Article
Real-World Effectiveness of Different Nutraceutical Formulations on Pain Intensity of Subjects with Diabetic Peripheral Neuropathy: An Observational, Retrospective, Case–Control Study
by Laura Armeli Grigio, Denisa Boci, Giacoma Di Vieste, Gianluca Cassanelli, Oscar Massimiano Epis, Alessandro Viadana, Federico Bertuzzi and Basilio Pintaudi
Biomedicines 2025, 13(6), 1407; https://doi.org/10.3390/biomedicines13061407 - 8 Jun 2025
Viewed by 775
Abstract
Background/Objectives. Diabetic peripheral neuropathy is a debilitating disease-related complication with a significant impact on quality of life. Its management represents a therapeutic challenge. Antioxidant agents such as α-lipoic acid, N-acetyl cysteine, and glutatione may be useful treatment strategies. Methods. A real-world, [...] Read more.
Background/Objectives. Diabetic peripheral neuropathy is a debilitating disease-related complication with a significant impact on quality of life. Its management represents a therapeutic challenge. Antioxidant agents such as α-lipoic acid, N-acetyl cysteine, and glutatione may be useful treatment strategies. Methods. A real-world, observational, retrospective, case–control study involving consecutive subjects with type 2 diabetes with diabetic peripheral neuropathy was performed. Participants who were supplemented with three different formulations for 12 weeks (high-dose α-lipoic acid (800 mg); low-dose α-lipoic acid (100 mg) plus glutathione (200 mg) plus Vitamin D (800 IU); N-acetyl cysteine (600 mg) plus glutathione (200 mg) plus Vitamin D (800 IU)) were compared with a non-treated control group. Questionnaires aimed at investigating the degree of disability and quality of life were administered. The primary endpoint was the change in neuropathic pain intensity measured by the Numerical Rating Scale (NRS). Results. Among 750 consecutive screened subjects with type 2 diabetes, 98 (13%) had diabetic neuropathy (mean age 66.7 ± 7.6 years, diabetes duration 11.3 ± 6.7 years, HbA1c 8.1 ± 1.5%, 43.8% insulin-treated). When comparing the differences between treatment groups in the changes in individual questionnaire scores between baseline and follow-up, all three supplements showed significant reductions compared to the control group in the NRS scale scores. No side effects have been reported during the study. Conclusions. As well as lipoic acid, other substances with specific activity on the genesis of neuropathic pain, such as N-acetyl cysteine and glutathione, have proved effective in reducing the intensity of pain. Full article
(This article belongs to the Special Issue Novel Biomarker and Treatments for Diabetic Neuropathy)
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20 pages, 6956 KiB  
Article
Chiral Growth of Gold Horns on Polyhedrons for SERS Identification of Enantiomers and Polarized Light-Induced Photothermal Sterilization
by Bowen Shang and Guijian Guan
Materials 2025, 18(11), 2627; https://doi.org/10.3390/ma18112627 - 4 Jun 2025
Viewed by 533
Abstract
The integration of chirality into nanomaterials holds significant potential for improving molecular recognition and biomedical technologies. In this work, we fabricated novel chiral horned gold nanostructures (HNS) by controlling the concentration of chiral ligands L-/D-cysteine (Cys). The unique three-dimensional morphology with horns-rotational arrangement [...] Read more.
The integration of chirality into nanomaterials holds significant potential for improving molecular recognition and biomedical technologies. In this work, we fabricated novel chiral horned gold nanostructures (HNS) by controlling the concentration of chiral ligands L-/D-cysteine (Cys). The unique three-dimensional morphology with horns-rotational arrangement enables synergistic optimization of chiral optical responses and surface-enhanced Raman scattering (SERS) performance. The proposed chiral HNSs can be used to recognize amino acid enantiomers, in which homochiral amino acid has distinct affinities to the chiral HNSs of homogeneous handedness. The 4-mercaptobenzoic acid (4-MPBA)-modified D-HNS demonstrates significantly enhanced targeting affinity for D-amino acids in the Escherichia coli (E. coli) cell wall, enabling successful amplification of SERS signals and advancing bacterial detection methodologies. By demonstrating the rotation-selective interaction between chiral HNSs and circularly polarized light (CPL), D-HNS exhibits excellent photothermal conversion efficiency under right-handed circularly polarized light (RCP) irradiation. This enables the synergistic combination of targeted physical disruption and photothermal sterilization, which leads to efficient eradication of E. coli. The D-HNS hydrogel composite system further expands the practical application of photothermal sterilization. Altogether, chiral HNSs have achieved SERS detection of bacteria and efficient polarization photothermal sterilization, which helps further develop applications based on chiral nanomaterials. Full article
(This article belongs to the Section Advanced Nanomaterials and Nanotechnology)
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16 pages, 1481 KiB  
Article
Arsenite-Mediated Transcriptional Regulation of Glutathione Synthesis in Mammalian Primary Cortical Astrocytes
by Jacob P. Leisawitz, Jiali He, Caroline Baggeroer and Sandra J. Hewett
Int. J. Mol. Sci. 2025, 26(11), 5375; https://doi.org/10.3390/ijms26115375 - 4 Jun 2025
Viewed by 580
Abstract
Arsenic, a potent metalloid contaminant of drinking water, is known for its ability to act as an initiator and modulator of disease in a variety of human tissues. Upon ingestion, arsenic is bio-transformed in the liver into a variety of metabolites, including arsenite. [...] Read more.
Arsenic, a potent metalloid contaminant of drinking water, is known for its ability to act as an initiator and modulator of disease in a variety of human tissues. Upon ingestion, arsenic is bio-transformed in the liver into a variety of metabolites, including arsenite. Arsenite permeates the blood–brain barrier (BBB), inducing oxidative stress that can be detrimental to brain neurons. As the primary glial cell at the BBB interface, astrocytes play a pivotal role in detoxifying xenobiotics such as arsenite via the production of the tripeptide antioxidant γ-glutamylcysteine, or glutathione (GSH). In this study, we assessed the mRNA levels of key components of the GSH synthetic pathway in astrocytes exposed to arsenite compared to vehicle controls. These components included xCT [substrate-specific light chain of the substrate importing transporter, system xc (Sxc)], glutamate-cysteine ligase [both catalytic (GCLC) and modifying (GCLM) subunits], and glutathione synthetase (GS). Additionally, we analyzed protein levels of some components by Western blotting and evaluated functional activity of Sxc using a fluorescence-based cystine uptake assay. Finally, we utilized a luminescence-based glutathione assay to determine the intracellular and extracellular GSH content in arsenite-treated cells. Arsenite significantly increased xCT, GCLC, GCLM, and GS mRNA levels, an effect blocked by the transcriptional inhibitor actinomycin D (ActD). A corresponding increase in Sxc activity was also observed in the arsenite treatment groups, along with significant increases in GCLC and GCLM protein expression. However, no increase in GS protein expression was detected. Finally, arsenite treatment significantly increased extracellular GSH levels, an effect which was also prevented by the inclusion of ActD. Overall, our study provides evidence that arsenite transcriptionally regulates several cellular processes necessary for GSH synthesis in primary cortical astrocyte cultures, thereby contributing to a better understanding of how this environmental toxicant influences antioxidant defenses in the brain. However, these results should be interpreted with caution regarding their applicability to vivo systems. Full article
(This article belongs to the Special Issue The Role of Glutathione Metabolism in Health and Disease)
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27 pages, 5400 KiB  
Article
Computational Modeling of Low-Abundance Proteins in Venom Gland Transcriptomes: Bothrops asper and Bothrops jararaca
by Joseph Espín-Angulo and Doris Vela
Toxins 2025, 17(6), 262; https://doi.org/10.3390/toxins17060262 - 22 May 2025
Viewed by 670
Abstract
Snake venoms contain numerous toxic proteins, but low-abundance proteins often remain uncharacterized due to identification challenges. This study employs a bioinformatics approach to identify and structurally model low-abundance proteins from the venom gland transcriptomes of Bothrops asper and Bothrops jararaca. Using tools [...] Read more.
Snake venoms contain numerous toxic proteins, but low-abundance proteins often remain uncharacterized due to identification challenges. This study employs a bioinformatics approach to identify and structurally model low-abundance proteins from the venom gland transcriptomes of Bothrops asper and Bothrops jararaca. Using tools such as tblastn, Jalview, and CHIMERA, we analyzed sequences and structural features of proteins including arylsulfatase, CRISP (Cysteine-Rich Secretory Protein), von Willebrand factor type D (vWFD), and dihydroorotate dehydrogenase (DHODH), and identified potential new isoforms of SVMP-PIIIb (Ba_1) and botrocetin in B. asper. Protein models were generated with AlphaFold2, compared with crystallized structures from the Protein Data Bank (PDB), and validated using Procheck, ERRAT, and Verify3D. Conserved motifs and domains were annotated through Pfam and InterPro, revealing structural elements that suggest possible roles in venom physiology and toxicity. These findings emphasize the potential of computational biology to characterize structurally relevant but experimentally inaccessible venom proteins, and to lay the groundwork for future functional validation. Full article
(This article belongs to the Special Issue Transcriptomic and Proteomic Study on Animal Venom: Looking Forward)
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16 pages, 2145 KiB  
Article
Impact of Combined Macronutrient Diet on Amino Acids and Amines Plasma Levels
by Celia Chicharro, Patricia Romero-Marco, Luz M. González, Laura González-Rodríguez, Laura Mateo-Vivaracho, Eva Guillamón, Francisco Miguel-Tobal, Fernando Bandrés, Guillermo Gervasini, Ana Fernández-Araque and Zoraida Verde
Nutrients 2025, 17(10), 1694; https://doi.org/10.3390/nu17101694 - 16 May 2025
Viewed by 552
Abstract
Background: Nutritional biomarkers are essential for assessing the impact of dietary interventions on health. Amino Acids (AA) serve as potential biomarkers due to their role in metabolic pathways, although the influence of combining macronutrients on AA metabolism remains unclear. Objective: This study aims [...] Read more.
Background: Nutritional biomarkers are essential for assessing the impact of dietary interventions on health. Amino Acids (AA) serve as potential biomarkers due to their role in metabolic pathways, although the influence of combining macronutrients on AA metabolism remains unclear. Objective: This study aims to evaluate the effects of combining macronutrients (fat, protein, and fiber) on AA metabolism. Methods: A dietary intervention was conducted with 41 participants who were assigned to two groups: the Protein Fat (PF) group and the Protein Fat and Fiber (PFF) group. AA concentration was measured using LC-MS/MS. Results: Our findings indicated that combining macronutrients reduced plasma levels of AA with statistically significant differences between the two diets (p < 0.05 for EAA and BCAA; p < 0,01 for NEAA and AAA). Metabolic profile analysis showed differences between the groups, especially at 55 days (55 D) (R2X: 0.749; R2Y: 0.589; Q2: 0.582, p < 0.001) and 98 days (98 D) (R2X: 0.886; R2Y: 0.7; Q2: 0.605, p < 0.001). Five metabolites (leucine, asparagine, alanine, cysteine, and creatine) were the most influential factors in group differentiation (VIP score), with values ranging between 1.04 and 1.75. Conclusions: The combination of macronutrients led to a significant reduction in plasma amino acids and amines in the PFF group, with recovery post-intervention. These findings highlight the possibility that these metabolites are related to different health statuses. Full article
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15 pages, 6161 KiB  
Article
Chiral-Dependent Redox Capacitive Biosensor Using Cu-Cys-GSH Nanoparticles for Ultrasensitive H2O2 Detection
by Duygu Yilmaz Aydin, Jie Jayne Wu and Jiangang Chen
Biosensors 2025, 15(5), 315; https://doi.org/10.3390/bios15050315 - 14 May 2025
Viewed by 488
Abstract
Copper-thiolate nanostructures, formed through the self-assembly of cysteine (Cys) and glutathione (GSH) with copper ions, offer a versatile platform for redox-active applications due to their structural stability and chemical functionality. In this study, Cu-Cys-GSH nanoparticles were synthesized and employed to develop a capacitive [...] Read more.
Copper-thiolate nanostructures, formed through the self-assembly of cysteine (Cys) and glutathione (GSH) with copper ions, offer a versatile platform for redox-active applications due to their structural stability and chemical functionality. In this study, Cu-Cys-GSH nanoparticles were synthesized and employed to develop a capacitive biosensor for the ultralow concentration detection of hydrogen peroxide (H2O2). The detection mechanism leverages a Fenton-like reaction, where H2O2 interacts with Cu-Cys-GSH nanoparticles to generate hydroxyl radicals (·OH) through redox cycling between Cu2+ and Cu+ ions. These redox processes induce changes in the sensor’s surface charge and dielectric properties, enabling highly sensitive capacitive sensing at gold interdigitated electrodes (IDEs). The influence of chirality on sensing performance was investigated by synthesizing nanoparticles with both L- and D-cysteine enantiomers. Comparative analysis revealed that the stereochemistry of cysteine impacts the catalytic activity and sensor response, with Cu-L-Cys-GSH nanoparticles exhibiting superior performance. Specifically, the biosensor achieved a linear detection range from 1.0 fM to 1.0 pM and demonstrated an ultra-sensitive detection limit of 21.8 aM, outperforming many existing methods for H2O2 detection. The sensor’s practical performance was further validated using milk and saliva samples, yielding high recovery rates and confirming its robustness and accuracy for real-world applications. This study offers a disposable, low-cost sensing platform compatible with sustainable healthcare practices and facilitates easy integration into point-of-care diagnostic systems. Full article
(This article belongs to the Special Issue Innovative Biosensing Technologies for Sustainable Healthcare)
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16 pages, 1717 KiB  
Article
Anti-Candida Activity of Cysteine-Modified Amidated Decoralin in the Presence of Engineered Nanomaterials
by Vânia Rocha, Helena Almeida, Bruno Sarmento and José das Neves
Pharmaceutics 2025, 17(4), 460; https://doi.org/10.3390/pharmaceutics17040460 - 2 Apr 2025
Cited by 1 | Viewed by 727
Abstract
Background: Candidiasis remains a chief concern in global healthcare. Drug safety issues and increasing resistance make it urgent to develop alternative antifungal agents, namely antimicrobial peptides. Amidated decoralin (Dec-CONH2) possesses considerable anti-Candida activity, and its association with nanocarriers could help [...] Read more.
Background: Candidiasis remains a chief concern in global healthcare. Drug safety issues and increasing resistance make it urgent to develop alternative antifungal agents, namely antimicrobial peptides. Amidated decoralin (Dec-CONH2) possesses considerable anti-Candida activity, and its association with nanocarriers could help in enhancing efficacy while reducing intrinsic toxicity to the host. Methods: We studied an N-terminal cysteine-modified version of the peptide (Cys-Dec-CONH2) and screened the effects of different nanosystems (polymeric nanoparticles (NPs), liposomes and gold NPs) on its activity against azole-sensitive and azole-resistant Candida species using a clinically relevant in vitro assay. Results: The antifungal activity of Cys-Dec-CONH2 was maintained (minimum inhibitory concentration (MIC) = 16–64 µg/mL), but the presence of poly(d,l-lactic-co-glycolic acid) (PLGA)- and polycaprolactone-based NPs impaired the antifungal effect of the peptide (MIC > 256 µg/mL). This effect was milder for polystyrene-based NPs, liposomes, and gold NPs (MIC ≤ 128 µg/mL). Additionally, the covalent surface functionalization of PLGA-based NPs with Cys-Dec-CONH2 or the presence of relevant biomolecules (albumin and mucin) resulted in complete inhibition of antifungal activity. Conclusions: Our data suggest that Cys-Dec-CONH2 is able to establish strong interfacial interactions with different nanomaterials, which need to be considered when developing nanomedicines based on this peptide for the management of candidiasis. Full article
(This article belongs to the Section Nanomedicine and Nanotechnology)
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16 pages, 3425 KiB  
Article
Circadian Regulation of Vitamin D Target Genes Reveals a Network Shaped by Individual Responsiveness
by Parcival Maissan and Carsten Carlberg
Nutrients 2025, 17(7), 1204; https://doi.org/10.3390/nu17071204 - 29 Mar 2025
Viewed by 1061
Abstract
Background: In humans, vitamin D3 synthesis follows a day–night rhythm due to its UV-B-dependent production. Results: As part of the VitDHiD intervention study, we identified 87 in vivo vitamin D target genes with circadian expression patterns in immune cells, forming a regulatory [...] Read more.
Background: In humans, vitamin D3 synthesis follows a day–night rhythm due to its UV-B-dependent production. Results: As part of the VitDHiD intervention study, we identified 87 in vivo vitamin D target genes with circadian expression patterns in immune cells, forming a regulatory network centered on transcription factors and membrane receptors. These genes exhibit a narrow basal expression range, with 80% downregulated upon vitamin D3 supplementation. Clustering analysis revealed six distinct gene groups, with the two most prominent clusters driven by the transcription factor CSRNP1 (cysteine- and serine-rich nuclear protein 1) and GAS7 (growth arrest-specific 7), a known differentiation inducer. Among the 25 VitDHiD study participants, we identified two subgroups distinguished by significant differences in the responsiveness of 14 in vivo vitamin D target genes. These genes encode transcription factors like CSRNP1, as well as metabolic enzymes and transporters, including NAMPT (nicotinamide phosphoribosyltransferase), PFKFB3 (6-phosphofructo-2-kinase/fructose-2,6-bisphosphatase 3), and SLC2A3 (solute carrier family 2 member 3). Notably, all 14 genes possess a vitamin D receptor-binding enhancer within a reasonable distance of their transcription start site. Conclusions: These findings highlight a novel link between vitamin D signaling and circadian gene regulation, with potential implications for personalized supplementation strategies. Full article
(This article belongs to the Section Nutrigenetics and Nutrigenomics)
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14 pages, 3075 KiB  
Article
Dynamic Interference Testing—Unexpected Results Obtained with the Abbott Libre 2 and Dexcom G6 Continuous Glucose Monitoring Devices
by Hendrick Jensch, Steven Setford, Nicole Thomé, Geethan Srikanthamoorthy, Lea Weingärtner, Mike Grady, Elizabeth Holt and Andreas Pfützner
Sensors 2025, 25(7), 1985; https://doi.org/10.3390/s25071985 - 22 Mar 2025
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Abstract
Background: Sensors for continuous glucose monitoring (CGM) are now commonly used by people with type 1 and type 2 diabetes. However, the response of these devices to potentially interfering nutritional, pharmaceutical, or endogenous substances is barely explored. We previously developed an in vitro [...] Read more.
Background: Sensors for continuous glucose monitoring (CGM) are now commonly used by people with type 1 and type 2 diabetes. However, the response of these devices to potentially interfering nutritional, pharmaceutical, or endogenous substances is barely explored. We previously developed an in vitro test method for continuous and dynamic CGM interference testing and herein explore the sensitivity of the Abbott Libre2 (L2) and Dexcom G6 (G6) sensors to a panel of 68 individual substances. Methods: In each interference experiment, L2 and G6 sensors were exposed in triplicate to substance gradients from zero to supraphysiological concentrations at a stable glucose concentration of 200 mg/dL. YSI Stat 2300 Plus was used as the glucose reference method. Interference was presumed if the CGM sensors showed a mean bias of at least ±10% from baseline with a tested substance at any given substance concentration. Results: Both L2 and G6 sensors showed interference with the following substances: dithiothreitol (maximal bias from baseline: L2/G6: +46%/−18%), galactose (>+100%/+17%), mannose (>+100%/+20%), and N-acetyl-cysteine (+11%/+18%). The following substances were found to interfere with L2 sensors only: ascorbic acid (+48%), ibuprofen (+14%), icodextrin (+10%), methyldopa (+16%), red wine (+12%), and xylose (>+100%). On the other hand, the following substances were found to interfere with G6 sensors only: acetaminophen (>+100%), ethyl alcohol (+12%), gentisic acid (+18%), hydroxyurea (>+100%), l-cysteine (−25%), l-Dopa (+11%), and uric acid (+33%). Additionally, G6 sensors could subsequently not be calibrated for use after exposure to dithiothreitol, gentisic acid, l-cysteine, and mesalazine (sensor fouling). Conclusions: Our standardized dynamic interference testing protocol identified several nutritional, pharmaceutical and endogenous substances that substantially influenced L2 and G6 sensor signals. Clinical trials are now necessary to investigate whether our findings are of relevance during routine care. Full article
(This article belongs to the Section Chemical Sensors)
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