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Search Results (2,749)

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25 pages, 3089 KiB  
Article
Microencapsulation and Probiotic Characterization of Lactiplantibacillus plantarum LM-20: Therapeutic Application in A Murine Model of Ulcerative Colitis
by Cynthia Garfias Noguez, Morayma Ramírez Damián, Alicia Ortiz Moreno, Yazmín Karina Márquez Flores, Liliana Alamilla Beltrán, Mario Márquez Lemus, Luis G. Bermúdez Humarán and María Elena Sánchez Pardo
Nutrients 2025, 17(5), 749; https://doi.org/10.3390/nu17050749 (registering DOI) - 20 Feb 2025
Abstract
Background: Microencapsulation improves the storage, handling, and administration of probiotics by protecting them from environmental factors and adverse conditions in the gastrointestinal tract. This process facilitates their controlled delivery in the body, which can simplify their use in therapies without compromising their therapeutic [...] Read more.
Background: Microencapsulation improves the storage, handling, and administration of probiotics by protecting them from environmental factors and adverse conditions in the gastrointestinal tract. This process facilitates their controlled delivery in the body, which can simplify their use in therapies without compromising their therapeutic efficacy. Objectives: This study investigates the microencapsulation of Lactiplantibacillus plantarum LM-20, its probiotic properties, and its effects in a murine model of ulcerative colitis. Methods/Results: Synbiotic microencapsulation was carried out using spray drying with maltodextrin, gum Arabic, and inulin, achieving an encapsulation efficiency of 90.76%. The resulting microcapsules exhibited remarkable resistance to simulated gastrointestinal conditions in vitro, maintaining a survival rate of 90%. The drying process did not compromise the probiotic characteristics of the bacteria, as they demonstrated enhanced auto-aggregation, hydrophobicity, and phenol tolerance. The therapeutic potential of the microencapsulated synbiotic was evaluated in a murine model of dextran sodium sulfate-induced ulcerative colitis. The results revealed that mice treated with microencapsulated Lactiplantibacillus plantarum LM-20 showed an 83.3% reduction in the disease activity index (DAI) compared to the ulcerative colitis control group. Moreover, a significant decrease was observed in pro-inflammatory cytokine levels (IL-1β and TNF-α) and myeloperoxidase activity, with values comparable to those of the healthy control group. Conclusions: These findings suggest that microencapsulated Lactiplantibacillus plantarum LM-20 could be a promising candidate for therapeutic applications in the prevention and management of ulcerative colitis. Full article
(This article belongs to the Section Prebiotics and Probiotics)
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Figure 1

Figure 1
<p>Representation of experiment duration and treatment times. Created with BioRender.com, 2024.</p>
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<p>Colony morphology on MRS agar and microscopic morphology of <span class="html-italic">Lactiplantibacillus plantarum</span> LM-20 using Gram staining identified under a 100× immersion objective optical microscope.</p>
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<p>Microscopic morphology of (<b>a</b>) SynLM20 and (<b>b</b>) SynLp115 synbiotic microcapsules and (<b>c</b>) microcapsules without bacteria (Microencapsulated inulin—MI) observed in Scanning Electron Microscopy (FEI-ThermoFisher Scientific, ESEM Quanta FEG 250, USA), under a voltage of 15 kV (kilovolts) and an increase of 5000× (amplification). The particle size was determined using the ImageJ program (version 1.53t) (National Institutes of Health, USA).</p>
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<p>Percentage of resistance to phenol of <span class="html-italic">Lactiplantibacillus plantarum</span> LM20 and Lp115, non-microencapsulated and microencapsulated in a synbiotic product by spray drying. Data are represented as mean ± standard error. Bars labeled with the same letters are not significantly different from each other.</p>
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<p>Percentage of hydrophobicity (<b>a</b>) and auto-aggregation (<b>b</b>) of <span class="html-italic">Lactiplantibacillus plantarum</span> LM20 and Lp115, non-microencapsulated and microencapsulated in a synbiotic product by spray drying. Data are represented as mean ± standard error. Bars labeled with the same letters are not significantly different from each other, while those labeled with different letters indicate significant differences from each other. In the case of hydrophobicity, different letters indicate significant differences <span class="html-italic">p</span> ≤ 0.0001. For auto-aggregation, the letters “a, b, e, f” indicate significant differences <span class="html-italic">p</span> ≤ 0.05, the letters “c, d” indicate significant differences <span class="html-italic">p</span> ≤ 0.01, and the letter “g” indicates that there is no significant difference.</p>
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<p>Evaluation of experimental animals’ disease activity index (DAI) during the trial period. From day 1 to day 7 (the day before the administration of DSS), no significant differences were observed, nor during the first 3 days after the administration of DSS. From day 12 onwards, significant differences were observed in the study groups, but for interesting and visual purposes, the results shown correspond to those obtained on the day of sacrifice, <span class="html-italic">n =</span> 12. The data are represented as a mean ± standard error. Letters indicate significant differences: bars labeled with the same letters are not significantly different from each other, while those labeled with different letters indicate a significant difference from each other.</p>
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<p>Evaluation of the weight–length ratio of the colon, <span class="html-italic">n</span> = 12. Data are represented as mean ± standard error. The letters indicate significant differences: bars labeled with the same letters are not significantly different from each other, while those labeled with different letters indicate significant differences from each other.</p>
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<p>Interleukin IL-1β colonic concentrations of mice with ulcerative colitis induced by DSS, <span class="html-italic">n =</span> 12. Data are represented as mean ± standard error. The letters indicate significant differences: bars labeled with the same letters are not significantly different from each other, while those labeled with different letters indicate significant differences from each other.</p>
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<p>TNF-α colonic concentrations of mice with ulcerative colitis induced by DSS, <span class="html-italic">n</span> = 12. Data are represented as mean ± standard error. The letters indicate significant differences: bars labeled with the same letters are not significantly different from each other, while those labeled with different letters indicate significant differences from each other.</p>
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<p>MPO colonic concentrations of mice with ulcerative colitis induced by DSS, <span class="html-italic">n</span> = 12. Data are represented as mean ± standard error. The letters indicate significant differences: bars labeled with the same letters are not significantly different from each other, while those labeled with different letters indicate significant differences from each other.</p>
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16 pages, 2591 KiB  
Article
Carbon Dioxide Selectivity over Ethane in Promising Bis Tri (Fluoromethylsulfonyl) Imide-Based Ionic Liquids
by Eric Quaye, Amr Henni and Ezeddin Shirif
Molecules 2025, 30(5), 984; https://doi.org/10.3390/molecules30050984 - 20 Feb 2025
Abstract
This research addresses the critical challenge of CO2 capture by exploring innovative ways to avoid ethane (C2H6) co-absorption in natural gas sweetening operations. The solubility of Ethane (C2H6) was measured in three ionic liquids [...] Read more.
This research addresses the critical challenge of CO2 capture by exploring innovative ways to avoid ethane (C2H6) co-absorption in natural gas sweetening operations. The solubility of Ethane (C2H6) was measured in three ionic liquids (ILs) with similar anions, 1-decyl-3-methyl imidazolium bis (trifluoro methylsulfonyl imide) [IL-1], 1-hexadecyl-3-methylimidazolium bis (trifluoro methylsulfonyl imide) [IL-2], and triethytetra-decyl ammonium bis (trifluoromethylsulfonyl imide) [IL-3]. The solubility experiments were investigated at 303.15 K and 343.15 K with pressures reaching 1.2 MPa. Among the ILs, [IL-2] exhibited the highest ethane absorption capacity due to its extended alkyl chain. The Peng-Robinson equation of state (PR-EoS) and three (3) distinct mixing rules provided robust correlations for the solubility data. Results demonstrate the inferior performance of [IL-1], [IL-2], and [IL-3] compared to Selexol/Genosorb 1753. The selectivity of Ethane (C2H6) over CO2 was determined, with the overall selectivity ranking as follows: [IL-1] > [IL-3] > [IL-2]. A comparison of these selectivity values with published IL data indicated that these three ILs are most effective when used in applications targeting CO2 capture in the absence of Ethane (C2H6), such as in the case of flue gas. They will most probably be used with an amine blend. Additionally, the Enthalpy and entropy of absorption provided valuable insights, demonstrating Ethane’s weaker interactions and lower solubility than CO2. These findings emphasize the critical role of IL structure in determining ethane solubility and highlight the potential of customized ILs for optimizing gas-separation processes. Full article
(This article belongs to the Section Molecular Liquids)
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<p>Ethane validation test with Florusse et al. [<a href="#B8-molecules-30-00984" class="html-bibr">8</a>] and Henni et al. [<a href="#B9-molecules-30-00984" class="html-bibr">9</a>] at 323.15 K.</p>
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<p>(<b>a</b>) C<sub>2</sub>H<sub>6</sub> absorption in IL-1; (<b>b</b>) C<sub>2</sub>H<sub>6</sub> absorption in IL-2; (<b>c</b>) C<sub>2</sub>H<sub>6</sub> solubility in IL-3.</p>
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<p>Henry’s Gas law constant (H) comparison for C<sub>2</sub>H<sub>6</sub> absorption between ILs in this study versus published ILs [<a href="#B9-molecules-30-00984" class="html-bibr">9</a>,<a href="#B12-molecules-30-00984" class="html-bibr">12</a>] and Selexol/Genesorb 1753 [<a href="#B10-molecules-30-00984" class="html-bibr">10</a>] at (<b>a</b>) 323.15 K and (<b>b</b>) 343.15 K.</p>
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<p>(<b>a</b>) Comparison to CO<sub>2</sub>/C<sub>2</sub>H<sub>6</sub> selectivity data for ILs published by Nath and Henni et al. [<a href="#B12-molecules-30-00984" class="html-bibr">12</a>] and Rayer al. [<a href="#B10-molecules-30-00984" class="html-bibr">10</a>] at 323 K; (<b>b</b>) at 343.15 K.</p>
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<p>Densities of the ILs used in this work.</p>
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27 pages, 3607 KiB  
Article
Predicting the Beneficial Effects of Cognitive Stimulation and Transcranial Direct Current Stimulation in Amnestic Mild Cognitive Impairment with Clinical, Inflammation, and Human Microglia Exposed to Serum as Potential Markers: A Double-Blind Placebo-Controlled Randomized Clinical Trial
by Ruth Alcalá-Lozano, Rocio Carmona-Hernández, Ana Gabriela Ocampo-Romero, Adriana Leticia Sosa-Millán, Erik Daniel Morelos-Santana, Diana Zapata Abarca, Dana Vianey Castro-de-Aquino, Edith Araceli Cabrera-Muñoz, Gerardo Bernabé Ramírez-Rodríguez, Ana Luisa Sosa Ortiz, Eduardo A. Garza-Villarreal, Ricardo Saracco-Alvarez and Jorge Julio González Olvera
Int. J. Mol. Sci. 2025, 26(4), 1754; https://doi.org/10.3390/ijms26041754 - 19 Feb 2025
Abstract
In amnestic mild cognitive impairment (aMCI), neuroinflammation evolves during disease progression, affecting microglial function and potentially accelerating the pathological process. Currently, no effective treatment exists, leading to explorations of various symptomatic approaches, though few target the underlying physiological mechanisms. Modulating inflammatory processes may [...] Read more.
In amnestic mild cognitive impairment (aMCI), neuroinflammation evolves during disease progression, affecting microglial function and potentially accelerating the pathological process. Currently, no effective treatment exists, leading to explorations of various symptomatic approaches, though few target the underlying physiological mechanisms. Modulating inflammatory processes may be critical in slowing disease progression. Cognitive stimulation (CS) and transcranial direct current stimulation (tDCS) applied to the left dorsolateral prefrontal cortex (l-DLPFC) show promise, but the results are heterogeneous. Thus, a randomized, double-blind, placebo-controlled clinical trial is currently underway. The first-stage results were examined after three weeks of intervention in two groups: active tDCS combined with CS and sham tDCS combined with CS. Twenty-two participants underwent two assessments: T0 (baseline) and T1 (after 15 sessions of tDCS, active or sham, and 9 sessions of CS). The results demonstrated that CS improved cognition, increased brain-derived neurotrophic factor (BDNF) levels, and reduced peripheral proinflammatory cytokine levels (interleukin IL-6 and chemokine CX3CL1) in serum. This decrease in IL-6 may promote microglial proliferation and survival as a modulatory effect response, while the increase in BDNF might suggest a regulatory mechanism in microglia–neuron interaction responses. However, tDCS did not enhance the cognitive or modulatory effects of CS, suggesting that longer interventions might be required to achieve substantial benefits. Full article
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<p>Clinical and Cognitive Parameters. Includes Montreal Cognitive Assessment Total (MoCA Total), Montreal Cognitive Assessment Memory Domain Score (MoCA Mem) and Montreal Cognitive Assessment Memory Index Score (MoCA MIS), Memory Failures of Everyday Questionnaire (MFEQ) and Clinical Global Impression-Severity (CGI-S) scores. Screening for Cognitive Impairment in Psychiatry (SCIP-S) includes Immediate Verbal Learning, Working Memory, Verbal Fluency, Delayed Verbal Learning, Speed of Processing and Total SCIP-S. ** <span class="html-italic">p</span> = 0.01; * <span class="html-italic">p</span> = 0.05.</p>
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<p>Biological parameters. (<b>A</b>) ELISA for interleukin-6 (IL-6), CX3CL1, and BDNF from the serum of patients with aMCI treated with active tDCS plus CS or sham tDCS plus CS. Protein quantifications (picograms/milliliter (pg/mL) and nanograms/mL (ng/mL)) were performed before (T0) and after (T1) the application of the interventions. Results are represented as the mean + error standard of the mean (SEM). (<b>B</b>) Experimental design for microglia cell culture. Microglia was cultured on 3 and 20% of O2 in the presence of fetal bovine serum (FBS) and penicillin/streptomycin (P/S). Thus, cells were plated on 96-well plates to be treated with serum of the participants with aMCI to measure cell proliferation and viability. A micrography of microglial cells is shown on panel (<b>B</b>). Scale bar = 30 μm. Charts show proliferation (left side) and viability (right side) of microglia cells cultured in 3 or 20% oxygen, respectively. *** <span class="html-italic">p</span> = 0.001; ** <span class="html-italic">p</span> = 0.01; * <span class="html-italic">p</span> = 0.05.</p>
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<p>Principal Component Analysis (PCA). PCA was applied to identify patterns of variance between the combined intervention (active-sham) and the times evaluated (T0–T1). The color dots represent the category (active or sham) to which the variables belong. The ellipses show clustering patterns while the arrows indicate the strength and direction. The variables C20, PRO20, and BDNF appear to be positively correlated since their vectors are close and point in the same direction (to the right). Variables whose vectors point in opposite directions are negatively correlated, i.e., as PRO20 and BDNF values increase, IL6 and CX3CL1 (FKN) values tend to decrease.</p>
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<p>The randomized clinical trial design in amnestic mild cognitive impairment participants (aMCI) consists of assessing participants before (T0) and after (T1) the interventions. The interventions include the administration of 15 sessions of active (active group) or sham (sham group) transcranial direct current stimulation (tDCS), five per week. Together with 9 sessions of cognitive stimulation (CS), three per week. Both interventions were distributed over three weeks.</p>
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16 pages, 2594 KiB  
Article
Study of the Viability of Separating Mixtures of Water–Bioethanol Using a Neoteric Solvent: 1-Decyl-3-methylimidazolium Bis(trifluoromethylsulfonyl)imide
by Maria-Pilar Cumplido, Javier de la Torre, Maria-Camila Arango, Josep Pasqual Cerisuelo and Amparo Chafer
Processes 2025, 13(2), 580; https://doi.org/10.3390/pr13020580 - 18 Feb 2025
Abstract
Following the successful utilization of various 1-alkyl-3-methylimidazolium bis(trifluoromethylsulfonyl)imide ionic liquids (ILs) as effective solvents in the extraction of ethanol, 1-propanol, and 2-propanol from water, we conducted experiments to determine the liquid–liquid equilibria data for the ternary mixture comprising water, ethanol, and 1-decyl-3-methylimidazolium bis(trifluoromethylsulfonyl)imide [...] Read more.
Following the successful utilization of various 1-alkyl-3-methylimidazolium bis(trifluoromethylsulfonyl)imide ionic liquids (ILs) as effective solvents in the extraction of ethanol, 1-propanol, and 2-propanol from water, we conducted experiments to determine the liquid–liquid equilibria data for the ternary mixture comprising water, ethanol, and 1-decyl-3-methylimidazolium bis(trifluoromethylsulfonyl)imide ([dmim][Tf2N]) at temperatures of 283.2 K, 303.2 K, and 323.2 K under atmospheric pressure. The thermodynamic parameters for both ternary mixtures were calculated using the non-random two-liquid (NRTL) and universal quasichemical (UNIQUAC) models, yielding favorable results across all investigated conditions (rmsd < 0.65%). Subsequently, we explored the efficiency of [dmim][Tf2N] in separating azeotropic mixtures by analyzing the distribution coefficient and selectivity (K2 and S greater than 1 in all cases, with maximum values of 3.551 and 10.878, respectively). Comparative assessments were made against the performance of various 1-alkyl-3-methylimidazolium bis(trifluoromethylsulfonyl)imide ILs and alcohols. The findings underscore the promising capabilities of [dmim][Tf2N] in achieving effective separation, providing valuable insights for potential applications in liquid–liquid extraction processes. Full article
(This article belongs to the Special Issue 1st SUSTENS Meeting: Advances in Sustainable Engineering Systems)
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<p>Liquid–liquid equilibria of water (1) + ethanol (2) + [dmim][Tf2N] (3) system at (a) T = 283.2 K. Experimental data: (•) aqueous phase, (○) [dmim][Tf2N] rich phase, (—) experimental tie lines, (˗ ˗ ˗) calculated tie line using UNIQUAC model. Binodal curve calculated using: (· · ·) NRTL model, (˗ ˗ ˗) UNIQUAC model.</p>
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<p>Liquid–liquid equilibria of water (1) + ethanol (2) + [dmim][Tf2N] (3) system at (a) T = 303.2 K. Experimental data: (•) aqueous phase, (○) [dmim][Tf2N] rich phase, (—) experimental tie lines, (˗ ˗ ˗) calculated tie line using UNIQUAC model. Binodal curve calculated using: (· · ·) NRTL model, (˗ ˗ ˗) UNIQUAC model.</p>
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<p>Liquid–liquid equilibria of water (1) + ethanol (2) + [dmim][Tf2N] (3) system at (a) T = 323.2 K. Experimental data: (•) aqueous phase, (○) [dmim][Tf2N] rich phase, (—) experimental tie lines, (˗ ˗ ˗) calculated tie line using UNIQUAC model. Binodal curve calculated using: (· · ·) NRTL model, (˗ ˗ ˗) UNIQUAC model.</p>
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<p>Influence of the temperature on liquid–liquid equilibrium of the water (1) + ethanol (2) + [dmim][Tf2N] (3) system. Experimental data: (•), at 283.2 K; (▲), at 303.2 K; (■), at 323.2 K. Comparison with literature data: (◦), at 283.2 K [<a href="#B50-processes-13-00580" class="html-bibr">50</a>].</p>
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<p>Influence of temperature on (<b>a</b>) distribution coefficient (the difference in solubility of ethanol in the two phases) and (<b>b</b>) selectivity (the capability of the IL to extract ethanol from the water) for the water (1) + ethanol (2) + [dmim][Tf2N] (3) system: (•), at 283.2 K; (▲), at 303.2 K; (■), at 323.2 K. (with lines).</p>
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<p>(<b>a</b>) Distribution coefficient (the difference in solubility of alcohol in the two phases) and (<b>b</b>) selectivity (the capability of the IL to extract alcohol from the water) for water (1) + alcohol (2) system at 323.2 K in different alcohols for [dmim][Tf2N] ionic liquid: (•), ethanol; (▲), 1-propanol [<a href="#B50-processes-13-00580" class="html-bibr">50</a>]; (■), 2-propanol [<a href="#B50-processes-13-00580" class="html-bibr">50</a>].</p>
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<p>(<b>a</b>) Distribution coefficient (the difference in solubility of ethanol in the two phases) and (<b>b</b>) selectivity (the capability of the IL to extract ethanol from the water) for water (1) + ethanol (2) system at 323.2 K in different ionic liquid: (▲), [emim][Tf2N] [<a href="#B45-processes-13-00580" class="html-bibr">45</a>]; (•), [bmim][Tf2N] [<a href="#B44-processes-13-00580" class="html-bibr">44</a>]; (♦), [hmim][Tf2N] [<a href="#B45-processes-13-00580" class="html-bibr">45</a>]; (■), [dmim][Tf2N].</p>
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22 pages, 4264 KiB  
Article
Seasonal Influences on Human Placental Transcriptomes Associated with Spontaneous Preterm Birth
by Khondoker M. Akram, Eleanor Dodd and Dilly O. C. Anumba
Cells 2025, 14(4), 303; https://doi.org/10.3390/cells14040303 - 18 Feb 2025
Abstract
Demographic studies have revealed a strong association between exposure to high ambient temperatures during pregnancy and increased risks of preterm birth (PTB). The mechanism underlying this association is unclear, but it is plausible that altered placental function may contribute to it. In this [...] Read more.
Demographic studies have revealed a strong association between exposure to high ambient temperatures during pregnancy and increased risks of preterm birth (PTB). The mechanism underlying this association is unclear, but it is plausible that altered placental function may contribute to it. In this study, we conducted differential gene expression analysis, gene set enrichment analysis (GSEA), and gene ontology (GO) analysis on bulk RNA-seq data from human placentas delivered at term and preterm during the warmer months compared to placentas delivered at term and preterm during the colder months in the UK. We detected 48 differentially expressed genes in preterm placentas delivered during the warmer months compared to preterm placentas delivered during the colder months, the majority of which were inflammatory cytokines and chemokines, including SERPINA1, IL1B, CCL3, CCL3L3, CCL4, CCL4L2, CCL20, and CXCL8. The GSEA positively enriched 17 signalling pathways, including the NF-κB, IL17, Toll-like receptor, and chemokine signalling pathways in preterm placentas delivered during warmer months. These results were not observed in the placentas delivered at term during the same times of the year. The GO analysis revealed several enhanced biological processes, including neutrophil, granulocyte, monocyte, and lymphocyte chemotaxis, as well as inflammatory and humoral immune responses in preterm placentas, but not in placentas delivered at term in the summer. We conclude that maternal exposure to warm environmental temperatures during pregnancy likely alters the placental transcriptomes towards inflammation and immune regulation, potentially leading to PTB. Full article
(This article belongs to the Special Issue Molecular Insight into the Pathogenesis of Spontaneous Preterm Birth)
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<p>Environmental temperature exposure during pregnancy. The rain-cloud plots showing monthly mean temperature (<b>A</b>) and average maximum temperature (<b>B</b>) when the women delivered. The clouds show the data kernel density, red dots with connectors show the mean, box plots show the median with the IQR, and each dot represents an individual subject. <span class="html-italic">n</span> = 10 warm group and <span class="html-italic">n</span> = 15 cold group (<b>A</b>,<b>B</b>). <span class="html-italic">p</span> values were calculated by the Mann–Whitney U test. (<b>C</b>) Box blots with jitters showing the duration of exposure of 4 groups of pregnant women to warm or cold weather temperatures prior to delivery. Data are presented as median values with the IQR, and each dot represents an individual subject. <span class="html-italic">p</span> = 0.82, as determined by one-way ANOVA.</p>
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<p>Differential gene expression analysis between warm and cold groups of placentas. (<b>A</b>) PCA plots showing group separation between warm and cold groups in preterm and term placentas. (<b>B</b>) Volcano plots showing DEGs in preterm-warm placentas compared to the preterm-cold group. (<b>C</b>) A heat map showing the expression of 48 significant DEGs in preterm-warm and preterm-cold placentas. (<b>D</b>) Signalling pathways in preterm-warm placentas positively enriched by GSEA. (<b>E</b>) The comparison of expression by a selected set of genes between term and preterm placentas.</p>
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<p>Cell-type enrichment and gene ontology analysis. (<b>A</b>) A bar plot showing the positive enrichment of cell types in preterm-warm placentas identified by GSEA. (<b>B</b>) A chord plot showing links between individual DEGs and enriched cell types. (<b>C</b>) A bar plot showing biological processes and cell components significantly enriched with the upregulated DEGs in preterm-warm placentas. (<b>D</b>) A bar plot showing disease phenotype enrichment with upregulated DEGs in the preterm-warm placentas.</p>
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<p>Differential gene expression and KEGG pathway analyses between preterm and term placentas. (<b>A</b>) A Venn diagram showing the overlap between significantly upregulated DEGs from the preterm-warm vs. preterm-cold and preterm-warm vs. term-warm group analyses. The common upregulated DEGs (FDR &lt; 0.05) are given in the box. (<b>B</b>) KEGG pathway enrichment with upregulated DEGs from preterm-warm vs. term-warm placentas. (<b>C</b>) Venn diagram showing the overlap of significantly enriched KEGG signalling pathways in Preterm-warm vs. Preterm-cold and Preterm-warm vs. Term-warm group analyses. The common upregulated pathways are shown in box (FDR &lt; 0.05).</p>
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<p>Individual gene expression in preterm placentas. (<b>A</b>–<b>L</b>) Box plots with jitters showing expressions of a selected set of DEGs in the warm and cold groups of preterm placentas. Data are presented as the median with the IQR. Each dot represents individual subjects. <span class="html-italic">p</span> values were determined by the Wilcoxon signed-rank test. (<b>M</b>) STRING protein–protein interaction analysis between the HPGD and related proteins (STRING v12).</p>
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<p>ELISA assay on placenta tissue. Box plots showing SERPINA1 protein concentrations in preterm-warm and preterm-cold placentas (<b>A</b>) and in term-warm and term-cold placentas (<b>B</b>). A protein concentration of 300 µg/mL was used as the loading concentration for each sample in the ELISA assay. Data are presented as median with interquartile range (IQR). Each dot represents an individual subject. The <span class="html-italic">p</span> value was determined by a two-tailed unpaired Student’s <span class="html-italic">t</span>-test. ns = not significant.</p>
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<p>Gene interaction network analysis by Cytoscape plug-in GeneMANIA. Striped bigger nodes indicate DEGs upregulated in preterm-warm placentas. Colour codes inside each node indicate their biological function as stated in the Functions legend. Coloured edges and their connections with other genes indicate the nature of interactions.</p>
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20 pages, 765 KiB  
Systematic Review
Translating Biomarker Research into Clinical Practice in Orthopaedic Trauma: A Systematic Review
by Alexander Baur and Augustine Mark Saiz
J. Clin. Med. 2025, 14(4), 1329; https://doi.org/10.3390/jcm14041329 - 17 Feb 2025
Abstract
Background/Objectives: Orthopaedic trauma management in polytrauma patients presents challenges, particularly in selecting between damage control orthopaedics (DCO) and early appropriate care (EAC). This systematic review evaluates these approaches and explores the role of biomarkers in optimising surgical timing. The primary objective of this [...] Read more.
Background/Objectives: Orthopaedic trauma management in polytrauma patients presents challenges, particularly in selecting between damage control orthopaedics (DCO) and early appropriate care (EAC). This systematic review evaluates these approaches and explores the role of biomarkers in optimising surgical timing. The primary objective of this review was to evaluate the potential clinical utility of biomarkers in guiding surgical timing and predicting perioperative complications. The secondary objective was to compare the effectiveness of DCO and EAC approaches, focusing on their impact on patient outcomes when controlled for Injury Severity Scores (ISSs). Methods: A systematic search of PubMed, MEDLINE, and Google Scholar identified studies focusing on fracture management (DCO versus EAC), timing protocols, and biomarkers in polytrauma patients. Twenty-seven studies met inclusion criteria. Results: Among the 27 studies, 12 evaluated biomarkers and 15 compared DCO and EAC. Point-of-care (POC) biomarkers, including lactate (p < 0.001; OR 1.305), monocyte L-selectin (p = 0.001; OR 1.5), and neutrophil L-selectin (p = 0.005; OR 1.56), demonstrated predictive value for sepsis, infection, and morbidity. CD16bright/CD62Ldim neutrophils were significant predictors of infection (p = 0.002). Advanced biomarkers, such as IL-6, IL-10, RNA IL-7R, HMGB1, and leptin offered prognostic insights but required longer processing times. No clear superiority was identified between DCO and EAC, with comparable outcomes when injury severity scores (ISS) were controlled. Conclusions: This systematic review highlights the challenge of translating biomarker research into clinical practice, identifying several point-of-care and advanced laboratory biomarkers with significant potential to predict complications like sepsis, infection, and MODS. Future efforts should focus on refining biomarker thresholds, advancing point-of-care technologies, and validating their role in improving surgical timing and trauma care outcomes. Full article
(This article belongs to the Special Issue Acute Trauma and Trauma Care in Orthopedics)
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<p>PRISMA flow diagram.</p>
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33 pages, 3219 KiB  
Article
Enhancing Energy Microgrid Sizing: A Multiyear Optimization Approach with Uncertainty Considerations for Optimal Design
by Sebastián F. Castellanos-Buitrago, Pablo Maya-Duque, Walter M. Villa-Acevedo, Nicolás Muñoz-Galeano and Jesús M. López-Lezama
Algorithms 2025, 18(2), 111; https://doi.org/10.3390/a18020111 - 17 Feb 2025
Abstract
This paper addresses the challenge of optimizing microgrid sizing to enhance reliability and efficiency in electrical energy supply. A comprehensive framework that integrates multiyear optimization with uncertainty considerations is presented to facilitate optimal microgrid design. The aim is to economically, safely, and reliably [...] Read more.
This paper addresses the challenge of optimizing microgrid sizing to enhance reliability and efficiency in electrical energy supply. A comprehensive framework that integrates multiyear optimization with uncertainty considerations is presented to facilitate optimal microgrid design. The aim is to economically, safely, and reliably supply electrical energy to communities with limited or no access to the main power grid, primarily utilizing renewable sources such as solar and wind technologies. The proposed framework incorporates environmental stochasticity, electrical demand uncertainty, and various electrical generation technologies. Electric power generation models are developed, and a metaheuristic optimization method is employed to minimize total costs while improving power supply reliability. The practical utility of the developed computational tool is emphasized, highlighting its significance in decision-making for microgrid installations. Utilizing real-world data, the approach involves a two-stage process: the first stage focuses on installation decisions, and the second evaluates operational performance using an iterated local search (ILS) optimization algorithm. Additionally, dispatch strategies are implemented to optimize computational time and enable real-time network modeling. The proposed microgrid sizing approach is a valuable asset for optimizing decision-making processes, significantly contributing to extending electricity coverage in non-interconnected zones while minimizing costs and ensuring steadfast reliability. Full article
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<p>Research Algorithm.</p>
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<p>Sample scenario: wind generation.</p>
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<p>Two-phase approach for the microgrid sizing problem.</p>
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<p>Diesel dispatch strategy.</p>
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<p>Diesel–renewable dispatch strategy.</p>
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<p>Battery–renewable dispatch strategy.</p>
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<p>Battery–renewable–diesel dispatch strategy.</p>
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<p>Complement 1.</p>
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<p>Two Stage ILS–optimization.</p>
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<p>Two Stage ILS–simulation with Dispatch Strategic.</p>
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<p>Comparison of different <math display="inline"><semantics> <mi>δ</mi> </semantics></math> values considering different limits of ENS.</p>
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<p>Comparison of different <math display="inline"><semantics> <mi>δ</mi> </semantics></math> values for different levels of allowed ENS: 1% Allowed ENS; 10% Allowed ENS.</p>
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<p>Variation 4—demand rises 10%.</p>
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<p>Variation 2–1.5% uncertainty in demand.</p>
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22 pages, 5559 KiB  
Article
Effect of Enzymatic Glycosylation on Film-Processing Properties and Biological Activities of Black Soybean Protein
by Yinglei Zhang, Xue Gong, Jing Wang, Boxin Dou, Lida Hou, Wei Xiao, Jiang Chang and Danting Li
Coatings 2025, 15(2), 238; https://doi.org/10.3390/coatings15020238 - 17 Feb 2025
Abstract
In this study, chitooligosaccharides (COS) were introduced into black soybean protein (BSP) using transglutaminase (TG) as a biocatalyst. The film-processing properties and physiological activities of the enzymatically glycosylated black soybean protein (EGBSP) were studied. The results showed that glycosylation decreased the surface hydrophobicity, [...] Read more.
In this study, chitooligosaccharides (COS) were introduced into black soybean protein (BSP) using transglutaminase (TG) as a biocatalyst. The film-processing properties and physiological activities of the enzymatically glycosylated black soybean protein (EGBSP) were studied. The results showed that glycosylation decreased the surface hydrophobicity, absolute value of the zeta potential, its minimum solubility, and film permeability of BSP by 69.86%, 6.04%, 36.68%, and 14.91%, respectively, while increasing the tensile strength and elongation at break of its protein film by 56.57% and 172.68%, respectively. The gel time was shortened, and the acid-induced gel properties of EGBSP were similar to those of BSP. The anticancer effect of EGBSP was evaluated by the tumor inhibition rate, flow cytometry, and morphology observation of an ascites tumor in H22 tumor-bearing mice. The immune organs (spleen, thymus), immune cells (lymphocytes, NK cells), and immune factors (IL-2, IL-12) of H22 tumor-bearing mice were detected to evaluate the immunomodulatory effects of EGBSP. The results showed that medium and high doses of BSP had positive effects on immune enhancement and anti-cancer activity of H22 tumor-bearing mice, while almost all doses of EGBSP showed significant effects. These results indicated that glycosylation significantly improved the anti-cancer effect and immunomodulatory activity of H22 tumor-bearing mice while prolonging their overall survival. In conclusion, the glycosylation method using microbial transglutaminase to catalyze the introduction of chitooligosaccharides into black bean protein can improve the film-processing properties and biological activities of BSP more effectively than the enzyme crosslinking method. Full article
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<p>Overall structure of microbial TGase (MTG): schematic ribbon drawing of the MTG molecule viewed from above the plate face. The secondary structure is numbered.</p>
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<p>Two typical reactions catalyzed by transglutaminase.</p>
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<p>Zeta potential of BSP and its modified products. Notes: black soybean protein isolated (BSP); enzymatic crosslinked black soybean protein isolated (ECBSP); enzymatically glycosylated black soybean protein (EGBSP). Different letters (a, b) indicate significant differences between groups (<span class="html-italic">p</span> &lt; 0.05).</p>
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<p>Solubility of BSP and its modified products in pH range of 2–11. Notes: black soybean protein isolated (BSP); enzymatic crosslinked black soybean protein isolated (ECBSP); enzymatically glycosylated black soybean protein isolated (EGBSP).</p>
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<p>Time sweep of GDL-induced gel process of BSP and its modified products’ dispersion solution. Notes: black soybean protein isolated (BSP); enzymatic crosslinked black soybean protein isolated (ECBSP); enzymatically glycosylated black soybean protein isolated (EGBSP).</p>
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<p>Effect of modification on TS value of BSP protein film. Notes: black soybean protein isolated (BSP); enzymatic crosslinked black soybean protein isolated (ECBSP); enzymatically glycosylated black soybean protein isolated (EGBSP). Different letters (a, b) indicate significant differences between groups (<span class="html-italic">p</span> &lt; 0.05).</p>
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<p>Effect of modification on E value of BSP protein film. Notes: black soybean protein isolated (BSP); enzymatic crosslinked black soybean protein isolated (ECBSP); enzymatically glycosylated black soybean protein isolated (EGBSP). Different letters (a–c) indicate significant differences between groups (<span class="html-italic">p</span> &lt; 0.05).</p>
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<p>Effect of modification on WVP value of BSP protein film. Notes: black soybean protein isolated (BSP); enzymatic crosslinked black soybean protein isolated (ECBSP); enzymatically glycosylated black soybean protein isolated (EGBSP). Different letters (a–c) indicate significant differences between groups (<span class="html-italic">p</span> &lt; 0.05).</p>
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<p>Effect of BSP and EGBSP on cellular morphology of mice with H<sub>22</sub> ascites tumor. Notes: black soybean protein isolated (BSP); enzymatically glycosylated black soybean protein isolated (EGBSP). Red arrows point to the cells with broken films.</p>
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<p>Effect of BSP and EGBSP on flow cytometry of mice with H<sub>22</sub> ascites tumor. (<b>a</b>) Normal control; (<b>b</b>) low dose of BSP; (<b>c</b>) middle dose of BSP; (<b>d</b>) high dose of BSP; (<b>e</b>) low dose of EGBSP; (<b>f</b>) middle dose of EGBSP; (<b>g</b>) high dose of EGBSP. Notes: black soybean protein isolated (BSP); enzymatically glycosylated black soybean protein isolated (EGBSP).</p>
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<p>Effect of BSP and EGBSP on flow cytometry of mice with H<sub>22</sub> ascites tumor. (<b>a</b>) Normal control; (<b>b</b>) low dose of BSP; (<b>c</b>) middle dose of BSP; (<b>d</b>) high dose of BSP; (<b>e</b>) low dose of EGBSP; (<b>f</b>) middle dose of EGBSP; (<b>g</b>) high dose of EGBSP. Notes: black soybean protein isolated (BSP); enzymatically glycosylated black soybean protein isolated (EGBSP).</p>
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<p>Effect of BSP and EGBSP on serum IL-2 of mice with tumor H<sub>22</sub>. Notes: black soybean protein isolated (BSP); enzymatically glycosylated black soybean protein isolated (EGBSP). Values are the mean ±SD for 10 mice. Different letters (a–c) indicate significant differences between groups (<span class="html-italic">p</span> &lt; 0.05).</p>
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<p>Effect of BSP and EGBSP on serum IL-12 of mice with tumor H<sub>22</sub>. Notes: black soybean protein isolated (BSP); enzymatically glycosylated black soybean protein isolated (EGBSP). Values are the mean ± SD for 10 mice. Different letters (a–c) indicate significant differences between groups (<span class="html-italic">p</span> &lt; 0.05).</p>
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21 pages, 8138 KiB  
Article
Assessing the Causal Relationship Between Plasma Proteins and Pulmonary Fibrosis: A Systematic Analysis Based on Mendelian Randomization
by Moxuan Han, Yan Cui, Zhengyuan Fang, He Li, Yueqi Wang, Mingwei Sima, Yan Bi and Donghui Yue
Biology 2025, 14(2), 200; https://doi.org/10.3390/biology14020200 - 14 Feb 2025
Abstract
Pulmonary fibrosis (PF) is a chronic interstitial lung disease characterized by the destruction of alveolar structures, the abnormal accumulation of extracellular matrix (ECM), and ultimately respiratory failure. Although previous studies have shown that plasma proteins play an important role in the onset and [...] Read more.
Pulmonary fibrosis (PF) is a chronic interstitial lung disease characterized by the destruction of alveolar structures, the abnormal accumulation of extracellular matrix (ECM), and ultimately respiratory failure. Although previous studies have shown that plasma proteins play an important role in the onset and progression of PF, there is currently a lack of systematic studies on causal relationships. To address the identified gap, the study employs the MR method to identify potential drug targets associated with PF. Plasma protein data (pQTL, exposure) were sourced from Ferkingstad et al. (n = 35,559), and PF-related summary statistics were obtained from the GWAS database (n = 469,126). The study integrates enrichment analysis, protein–protein interaction (PPI) networks, drug prediction, molecular docking, and single-cell sequencing to further evaluate the biological functions and pharmacological potential of the identified targets. In the MR analysis, 64 genetic loci were significantly associated with the occurrence of PF. Further reverse Mendelian analysis revealed a positive causal relationship between PF and genes such as NPTX1, IL31, and CTSE, suggesting that these proteins may play a promotive role in the onset and progression of pulmonary fibrosis. The PPI network analysis identified core genes such as CDH1, CRP, VTN, COL1A1, and MAPK8, which are involved in the key pathological processes of PF, including cell signaling, ECM remodeling, and immune responses. The drug prediction analysis identified potential drugs such as sorafenib, vitamin C, and vitamin E, and the molecular docking analysis showed good binding between the drugs and the proteins. The single-cell sequencing results showed that core genes were highly expressed in fibroblasts and alveolar type II cells, confirming their potential role in the pathogenesis of PF. This study successfully identifies 64 potential drug targets for PF, with 10 core targets considered particularly promising for clinical trials. These findings offer valuable insights into the molecular mechanisms underlying PF and open new avenues for the development of targeted therapies. This research may accelerate the development of effective PF treatments and reduce drug development costs by providing more precise and personalized approaches to managing the disease. Full article
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<p>Overview of the study design [<a href="#B13-biology-14-00200" class="html-bibr">13</a>].</p>
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<p>MR analysis results of pQTL on PF.</p>
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<p>GO enrichment results for three terms.</p>
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<p>KEGG enrichment results.</p>
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<p>PPI network construction diagram. (<b>A</b>): PPI network built with STRING. (<b>B</b>): Full PPI network of selected genes. Key clusters with hub genes highlighted in red. (<b>C</b>): Core sub-network showing interactions among top hub genes.</p>
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<p>Bar chart of drug prediction results.</p>
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<p>Gene–drug interaction network diagram.</p>
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<p>Docking results of available proteins small molecules.</p>
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<p>(<b>A</b>): t-SNE cell distribution plot. Based on single-cell RNA sequencing data, the t-SNE dimensionality reduction plot illustrates the heterogeneity of proximal airway stromal cells in the lung. Different colors represent cell subpopulations. (<b>B</b>): Key information of 22 cell clusters.</p>
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<p>Scatterplots showing the expression distribution of core genes across clusters in lung proximal airway stromal cells.</p>
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17 pages, 3264 KiB  
Article
Mitigating Post-Subarachnoid Hemorrhage Complications: Anti-Inflammatory and Anti-Apoptotic Effects of Anakinra in an Experimental Study
by Güven Kılıç, Berk Enes Engin, Amir Halabi, Cengiz Tuncer, Mehmet Ali Sungur, Merve Alpay, Adem Kurtuluş, Hakan Soylu, Ali Gök and Ömer Polat
J. Clin. Med. 2025, 14(4), 1253; https://doi.org/10.3390/jcm14041253 - 14 Feb 2025
Abstract
Background: Subarachnoid hemorrhage (SAH) is a severe neurological condition with high mortality and morbidity rates, often exacerbated by secondary complications such as inflammation, cerebral vasospasm, and apoptosis. Proinflammatory cytokines, including interleukin-1 (IL-1), tumor necrosis factor-alpha (TNF-α), and interleukin-6 (IL-6), play critical roles [...] Read more.
Background: Subarachnoid hemorrhage (SAH) is a severe neurological condition with high mortality and morbidity rates, often exacerbated by secondary complications such as inflammation, cerebral vasospasm, and apoptosis. Proinflammatory cytokines, including interleukin-1 (IL-1), tumor necrosis factor-alpha (TNF-α), and interleukin-6 (IL-6), play critical roles in these pathological processes. Anakinra, an IL-1 receptor antagonist, has demonstrated significant anti-inflammatory effects in various disease models. This study aimed to evaluate the efficacy of anakinra in mitigating inflammation, vasospasm, and apoptosis in an experimental rat model of SAH. Methods: Thirty-two male Sprague Dawley rats were divided into four groups: Control (healthy), SAH (no treatment), Saline (0.2 mL saline subcutaneously), and Anakinra (50 mg/kg subcutaneously, twice daily). Proinflammatory markers (CRP, TNF-α, IL-1, IL-6, and fibrinogen) were measured in serum and cerebrospinal fluid (CSF) at 3, 7, and 10 days post-SAH. Basilar artery diameter was evaluated histopathologically, and Caspase-3 expression was assessed immunohistochemically to determine apoptotic activity. Results: SAH significantly increased levels of CRP, TNF-α, IL-1, IL-6, and fibrinogen in both serum and CSF, reduced basilar artery diameter, and elevated Caspase-3 expression compared to the Control group. Saline treatment provided limited improvements, with inflammatory markers and histopathological parameters remaining elevated. Anakinra treatment significantly reduced inflammatory markers, restored basilar artery diameter, and lowered Caspase-3 expression, highlighting its efficacy in mitigating inflammation, vasospasm, and apoptosis. Conclusions: Anakinra effectively suppresses inflammation, alleviates cerebral vasospasm, and inhibits apoptosis in an experimental model of SAH. These findings suggest its potential as a therapeutic agent for managing SAH and its complications. Further research is needed to explore its clinical applicability and long-term effects. Full article
(This article belongs to the Section Clinical Neurology)
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<p>Representative photographs of hematoxylin and eosin staining. Objective magnification is 40× and scale bar is 20 µm.</p>
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<p>Representative photographs of cleaved Caspase-3 immunohistochemistry results. Objective magnification is 40× and scale bar is 20 µm.</p>
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<p>A comparison of (<b>a</b>) serum and (<b>b</b>) CSF CRP levels between groups. CRP values, which were statistically significantly increased in both serum and CSF with SAH compared to the Control group, decreased in the ANA treatment group. No significant difference was seen between the SAH and Saline groups in either serum or in CSF for the entire period of the experiment, and both groups showed significantly higher values compared to the Control group for all three periods of the experiment. While the decrease over the 3-day treatment period in the ANA group was not significant compared to the SAH and Saline groups, the levels of decrease were statistically significant over the 7-day and 10-day treatment periods both in serum and CSF. However, there was no statistically significant difference between the 3-day, 7-day, and 10-day treatment periods in the ANA group, although the CRP values continued to decrease over the treatment period and the lowest value that was closest to that of the Control group was detected on day 10. SAH: subarachnoid hemorrhage; ANA: anakinra; CSF: cerebrospinal fluid; CRP: C-reactive protein.</p>
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<p>A comparison of (<b>a</b>) serum and (<b>b</b>) CSF TNF-α levels between groups. TNF-α values, which were statistically significantly increased in both serum and CSF with SAH compared to the Control group, decreased in the ANA treatment group. No significant difference was seen between the SAH and Saline groups in either serum or in CSF for the entire period of the experiment, and both groups showed significantly higher values compared to the Control group for all three periods of the experiment. While the decrease over the 3-day treatment period in the ANA group was significant compared to the SAH and Saline groups in both serum and CSF, the decrease over the 7-day treatment period did not show a significant difference, although the measured levels of TNF-α in serum were similar. Again, the level of decrease in the ANA group was statistically significant over the 10-day treatment period in both serum and CSF. While there was no statistically significant difference in the ANA group among the 3-day, 7-day, and 10-day values in serum, a significant change was observed in CSF, revealing significantly lower levels of TNF-α for the 7-day and 10-day periods in comparison to the 3-day period. SAH: subarachnoid hemorrhage; ANA: anakinra; CSF: cerebrospinal fluid; TNF-α: tumor necrosis factor-alpha.</p>
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<p>A comparison of (<b>a</b>) serum and (<b>b</b>) CSF IL-1 levels between groups. IL-1 values, which were statistically significantly increased in both serum and CSF with SAH compared to the Control group, decreased in the ANA treatment group. No significant difference was seen between the SAH and Saline groups in either serum or in CSF for the entire period of the experiment, and both groups showed significantly higher values compared to the Control group for all three periods of the experiment. The levels of decrease in the ANA group for all three treatment periods were statistically significant both in serum and CSF when compared to the SAH and Saline groups. On the other hand, a significant change was observed among the 3-day, 7-day, and 10-day values in both serum and CSF, revealing significantly lower levels of IL-1 for the 10-day period in comparison to the 3-day and 7-day periods. SAH: subarachnoid hemorrhage; ANA: anakinra; CSF: cerebrospinal fluid; IL-1: interleukin 1.</p>
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<p>A comparison of (<b>a</b>) serum and (<b>b</b>) CSF IL-6 levels between groups. IL-6 values, which were statistically significantly increased in both serum and CSF with SAH compared to the Control group, decreased in the ANA treatment group. No significant difference was seen between the SAH and Saline groups in either serum or in CSF for the entire period of the experiment, and both groups showed significantly higher values compared to the Control group for all three periods of the experiment. While the decrease over the 3-day treatment period in the ANA group was not significant compared to the SAH and Saline groups, the levels of decrease were statistically significant over the 7-day and 10-day treatment periods in serum. In addition, there was no significant difference in CSF IL-6 levels between the ANA, SAH and Saline groups for any of the three treatment periods. While a significant change was observed in serum IL-6 in the ANA group among the 3-day, 7-day, and 10-day values, revealing significantly lower levels of IL-6 for the 7-day and 10-day periods in comparison to the 3-day period, there was no statistically significant difference in the ANA group among the 3-day, 7-day, and 10-day values in CSF. SAH: subarachnoid hemorrhage; ANA: anakinra; CSF: cerebrospinal fluid; IL-6: interleukin 6.</p>
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<p>A comparison of (<b>a</b>) serum and (<b>b</b>) CSF FIB levels between groups. FIB values, which were statistically significantly increased in serum with SAH compared to Control group, remained at similar levels in the ANA treatment group. No significant difference was seen between the ANA, SAH, and Saline groups in either serum or CSF for all periods of the experiment, except for the 10-day period, where only the ANA group was significantly different from the SAH and Saline groups and similar to the Control group. However, there was no statistically significant difference between the 3-day, 7-day, and 10-day treatment periods in the ANA group, although the FIB values continued to decrease over the treatment period and the lowest value that was closest to that of the control group was detected on day 10. SAH: subarachnoid hemorrhage; ANA: anakinra; CSF: cerebrospinal fluid; FIB: fibrinogen.</p>
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<p>A comparison of lumen diameters between groups. Lumen diameter, which was statistically significantly decreased with SAH compared to the Control group, increased significantly in the ANA treatment group. No significant difference was seen between the SAH and Saline groups, and both groups showed significantly lower values compared to the Control group. SAH: subarachnoid hemorrhage; ANA: anakinra; <span class="html-italic">p</span> values of multiple comparisons: control vs. SAH (<span class="html-italic">p</span> &lt; 0.001), control vs. saline (<span class="html-italic">p</span> &lt; 0.001), control vs. ANA (<span class="html-italic">p</span> = 0.002), SAH vs. saline (<span class="html-italic">p</span> = 0.999), SAH vs. ANA (<span class="html-italic">p</span> &lt; 0.001), and saline vs. ANA (<span class="html-italic">p</span> &lt; 0.001).</p>
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<p>A comparison of Caspase levels between groups. Caspase level, which was statistically significantly increased with SAH compared to the Control group, decreased significantly in the ANA treatment group. No significant difference was seen between the SAH and Saline groups, and both groups showed significantly lower values compared to the Control group. SAH: subarachnoid hemorrhage; ANA: anakinra; <span class="html-italic">p</span> values of multiple comparisons: control vs. SAH (<span class="html-italic">p</span> &lt; 0.001), control vs. saline (<span class="html-italic">p</span> &lt; 0.001), control vs. ANA (<span class="html-italic">p</span> = 0.001), SAH vs. saline (<span class="html-italic">p</span> = 0.984), SAH vs. ANA (<span class="html-italic">p</span> &lt; 0.001), and saline vs. ANA (<span class="html-italic">p</span> = 0.001).</p>
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22 pages, 12777 KiB  
Article
Effect of Food Matrix on Regulation of Intestinal Barrier and Microbiota Homeostasis by Polysaccharides Sulfated Carrageenan
by Xuke Shang, Juanjuan Guo and Peilin Chen
Foods 2025, 14(4), 635; https://doi.org/10.3390/foods14040635 - 14 Feb 2025
Abstract
Carrageenan (CGN) has side effects on the intestinal barrier. Damage to the intestinal barrier is associated with exposure to sulfate groups. Food matrix has significant influence on the exposure quantity of sulfate groups and conformation in κ-CGN, but the corresponding side effects are [...] Read more.
Carrageenan (CGN) has side effects on the intestinal barrier. Damage to the intestinal barrier is associated with exposure to sulfate groups. Food matrix has significant influence on the exposure quantity of sulfate groups and conformation in κ-CGN, but the corresponding side effects are not reported specifically. This study aimed to explore the regulatory effect of κ-CGN dissolved in aqueous (κ-CGN) and in 3% casein (κ-carrageenan-casein, κ-CC) on the intestinal barrier and microbiota homeostasis. Research has shown that both κ-CGN and κ-CC can induce different extents of intestinal barrier damage through disrupting microbiota homeostasis. Importantly, κ-CGN in casein with lower sulfate groups content was found to repair the intestinal barrier injury induced by an equivalent dose of κ-CGN aqueous through increasing the abundance of Oscillibacter and decreasing Weissella. These alleviating effects were reflected in lower levels of tumor necrosis factor (TNF)-α and C-reaction protein (CRP), higher levels of interleukin (IL)-10, raised secretion of mucus and goblet cells, and improved expression of epithelial cell compact proteins zonula occluden (ZO)-1 and mucin protein 2 (MUC2). This study states that κ-CGN in casein has a positive regulatory effect on the intestinal barrier damage compared to in aqueous solution, which can provide guidance for processing and utilization of CGN. Full article
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<p>The conformational characterizations of the κ-CGN and κ-CC. (<b>A</b>) The basic unit structures of κ-CGN. (<b>B</b>) Physical diagram and conformational characterizations of the κ-CGN in the simulated intestinal phase. (<b>C</b>) Physical diagram and conformational characterizations of the κ-CC in the simulated intestinal phase. (<b>D</b>) Confocal laser scanning microscopy images. The arrows indicate the microscopic morphological features of the sample observed under laser confocal microscopy. κ-CGN appears green while casein is red/orange.</p>
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<p>Effect of κ-CGN and κ-CC on colitis in mice (n = 8 for each group). (<b>A</b>) Body weight change from week 1 to 8. (<b>B</b>) Body weight change at week 8. (<b>C</b>) Fecal condition. (<b>D</b>) DAI scores change from week 1 to 8. (<b>E</b>) DAI scores change at week 8. (<b>F</b>) Spleen changes. (<b>G</b>) Spleen organ index at week 8. (<b>H</b>) Colon length change at week 8. (<b>I</b>) Colon condition. The arrows indicate the length of the colon, and the circles indicate the occurrence of congestion. Independent samples <span class="html-italic">t</span>-tests were used for a single comparison of differences between groups and multiple comparisons were performed using the Turkey post hoc test after a significant one-way ANOVA (<span class="html-italic">p</span> &lt; 0.05). Uppercase letters represent differences within the κ-CGN groups, lowercase letters represent differences within the κ-CC groups. “*” represents differences between the κ-CGN and κ-CC groups (<span class="html-italic">p</span> &lt; 0.05) and “**” represents differences between the κ-CGN and κ-CC groups (<span class="html-italic">p</span> &lt; 0.01).</p>
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<p>Effects of κ-CGN and κ-CC on inflammatory cytokines and the intestinal barrier (n = 8 for each group). (<b>A</b>–<b>C</b>) Serum inflammatory cytokines levels of TNF-α, CRP, and IL-10. (<b>D</b>) Images of HE staining. The dotted line indicates the surface of the irregular crypt and arrows indicate infiltration of inflammatory cells. (<b>E</b>) HAI scores. (<b>F</b>) Quantification of mucus secretion. (<b>G</b>) Images of AB-PAS staining. Circles and arrows indicate acidic mucus staining. Independent samples <span class="html-italic">t</span>-tests were used for a single comparison of differences between groups and multiple comparisons were performed using the Turkey post hoc test after a significant one-way ANOVA (<span class="html-italic">p</span> &lt; 0.05). Uppercase letters represent differences within the κ-CGN groups, lowercase letters represent differences within the κ-CC groups. “*” represents differences between the κ-CGN and κ-CC groups (<span class="html-italic">p</span> &lt; 0.05) and “**” represents differences between the κ-CGN and κ-CC groups (<span class="html-italic">p</span> &lt; 0.01). scale bar = 50 μm.</p>
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<p>Effect of κ-CC and κ-CGN on the expression of ZO-1 and MUC2 in mice (n = 8 for each group). (<b>A</b>,<b>B</b>) The mRNA levels of ZO-1 and MUC2. (<b>C</b>,<b>D</b>) Western bolt results and the protein expression of ZO-1. (<b>E</b>) The protein expression of MUC2. (<b>F</b>) Immunohistochemistry staining. The arrows indicated MUC2 staining. Independent samples <span class="html-italic">t</span>-tests were used for a single comparison of differences between groups and multiple comparisons were performed using the Turkey post hoc test after a significant one-way ANOVA (<span class="html-italic">p</span> &lt; 0.05). Uppercase letters represent differences within the κ-CGN groups, lowercase letters represent differences within the κ-CC groups. “*” represents differences between the κ-CGN and κ-CC groups (<span class="html-italic">p</span> &lt; 0.05) and “**” represents differences between the κ-CGN and κ-CC groups (<span class="html-italic">p</span> &lt; 0.01). scale bar = 50 μm.</p>
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<p>Effect of κ-CGN and κ-CC on the gut microbiota at genus (n = 8 for each group). (<b>A</b>) Alpha diversity. (<b>B</b>) Beta diversity. (<b>C</b>) Stacked column plot of microbial genus relative abundance. (<b>D</b>) Heatmap analysis of relative abundance of top 50 genera. (<b>E</b>) The Kruskal–Wallis test results for comparison of microbial abundance among six groups. (<b>F</b>) Relative abundance of differential bacteria in κ-CGN and κ-CC groups, which were calculated by a Wilcoxon rank sum test, * <span class="html-italic">p</span> &lt; 0.05, and ** <span class="html-italic">p</span> &lt; 0.01.</p>
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<p>LEfSe analysis of gut microbiota and Spearman’s analysis between the microbiota and biochemical indexes. (<b>A</b>) Taxonomic cladogram obtained from LEfSe analysis among six groups. Different colors indicate the enrichment of the biomarker taxa. The circle from inside to outside means the rank from kingdom to genus, and the circle size represents the taxa abundance in the community. (<b>B</b>) Circle bar of LDA scores from LEfSe analysis at genus (LDA &gt; 3). (<b>C</b>) Correlation analysis of characteristic microbiota and biochemical indexes in the κ-CGN group. (<b>D</b>) Correlation analysis of characteristic microbiota and biochemical indexes in the κ-CC group. The color scale represents the strength of correlation, ranging from 0.5 (strong positive correlation) to − 0.5 (strong negative correlation). * <span class="html-italic">p</span> &lt; 0.05, ** <span class="html-italic">p</span> &lt; 0.01.</p>
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<p>Schematic diagram of κ-CGN solution and κ-CC causing microbiota changes in mice. (The arrows indicate upward and downward changes in microbiota or physiological indicators.)</p>
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27 pages, 6858 KiB  
Article
Biofunctionalization of Collagen Barrier Membranes with Bone-Conditioned Medium, as a Natural Source of Growth Factors, Enhances Osteoblastic Cell Behavior
by Harshitha Ashoka Sreeja, Emilio Couso-Queiruga, Clemens Raabe, Vivianne Chappuis and Maria B. Asparuhova
Int. J. Mol. Sci. 2025, 26(4), 1610; https://doi.org/10.3390/ijms26041610 - 13 Feb 2025
Abstract
A key principle of guided bone regeneration (GBR) is the use of a barrier membrane to prevent cells from non-osteogenic tissues from interfering with bone regeneration in patients with hard tissue deficiencies. The aim of the study was to investigate whether the osteoinductive [...] Read more.
A key principle of guided bone regeneration (GBR) is the use of a barrier membrane to prevent cells from non-osteogenic tissues from interfering with bone regeneration in patients with hard tissue deficiencies. The aim of the study was to investigate whether the osteoinductive properties of bone-conditioned medium (BCM) obtained from cortical bone chips harvested at the surgical site can be transferred to a native bilayer collagen membrane (nbCM). BCM extracted within 20 or 40 min, which corresponds to a typical implant surgical procedure, and BCM extracted within 24 h, which corresponds to BCM released from the autologous bone chips in situ, contained significant and comparable amounts of TGF-β1, IGF-1, FGF-2, VEGF-A, and IL-11. Significant (p < 0.001) quantities of BMP-2 were only detected in the 24-h BCM preparation. The bioactive substances contained in the BCM were adsorbed to the nbCMs with almost 100% efficiency. A fast but sequential release of all investigated proteins occurred within 6–72 h, reflecting their stepwise involvement in the natural regeneration process. BCM-coated nbCM significantly (p < 0.05) increased the migratory, adhesive, and proliferative capacity of primary human bone-derived cells (hBC), primary human periodontal ligament cells (hPDLC), and an osteosarcoma-derived osteoblastic cell line (MG-63) compared to cells cultured on BCM-free nbCM. The high proliferative rates of MG-63 cells cultured on BCM-free nbCM were not further potentiated by BCM, indicating that BCM-coated nbCM has no detrimental effects on cancer cell growth. BCM-coated nbCM caused significant (p < 0.05) induction of early osteogenic marker gene expression and alkaline phosphatase activity, suggesting an important role of BCM-functionalized nbCM in the initiation of osteogenesis. The 24-h BCM loaded on the nbCM was the only BCM preparation that caused significant induction of late osteogenic marker gene expression. Altogether, our data define the pre-activation of collagen membranes with short-term-extracted BCM as a potential superior modality for treating hard tissue deficiencies via GBR. Full article
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Figure 1
<p>Release of various growth factors and cytokines involved in bone metabolism from cortical bone. Enzyme-linked immunosorbent assay (ELISA) quantification of TGF-β1, IGF-1, FGF-2, VEGF-A, IL-11, and BMP-2 proteins contained in bone-conditioned medium (BCM) extracted from cortical bone chips with Ringer’s solution (RS). BCM was collected at 20 min, 40 min, and 24 h. RS not containing bone particles represents the control (Ctrl) and exhibits no detectable levels of the proteins tested. Means ± SD from four independent BCM preparations from each type and significant differences to the control, *** <span class="html-italic">p</span> &lt; 0.001, ** <span class="html-italic">p</span> &lt; 0.01, are shown.</p>
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<p>Adsorption and release of TGF-β1 (<b>a</b>), IGF-1 (<b>b</b>), FGF-2 (<b>c</b>), VEGF-A (<b>d</b>), IL-11 (<b>e</b>), and BMP-2 (<b>f</b>) from nbCM coated with either the respective recombinant (r) protein or BCM preparation. The nbCM was incubated for 10 min at room temperature in either RS containing the respective recombinant protein at the average concentration measured in the BCM preparations (cf. <a href="#ijms-26-01610-f001" class="html-fig">Figure 1</a>) or each of the three BCM (20 min, 40 min, and 24 h). Hydration of the nbCM with RS was used as a control (Ctrl). Protein quantifications were performed by using colorimetric ELISA assays. Tables (a–f) represent: (1) quantifications of adsorbed protein (in percent); (2) total protein release (expressed as percent of adsorbed protein) from nbCM for a 4-day period; (3) the time point at which the highest protein release was observed (peak of release); (4) ELISA quantifications of the protein released before (and including) the peak expressed as percent of the total protein release for the entire test period (taken as 100%); (5) ELISA quantifications of the protein released after the peak until day 4 and expressed as in (4). Means ± SD from three independent experiments and significant differences (<span class="html-italic">p</span> &lt; 0.05) between the experimental groups are shown. Significance was indicated with the following symbols: # denotes significantly higher than recombinant protein; <span>$</span> denotes significantly higher than 20 min BCM; † denotes significantly higher than 40 min BCM; ‡ denotes significantly higher than 24 h BCM. Graphs represent the results from ELISA quantifications of the proteins measured in conditioned RS collected from the nbCM at the indicated time points over a 4-day period. Data represent means ± SD from three independent experiments. Significant differences between experimental groups, *** <span class="html-italic">p</span> &lt; 0.001, ** <span class="html-italic">p</span> &lt; 0.01, * <span class="html-italic">p</span> &lt; 0.05.</p>
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<p>Induced migration capacity of osteoblast-like cell lines toward BCM-coated nbCM. Migration of primary human bone-derived cells (hBC) (<b>a</b>,<b>b</b>), primary human periodontal ligament cells (hPDLC) (<b>c</b>,<b>d</b>), and an osteosarcoma-derived immortal cell line (MG-63) (<b>e</b>,<b>f</b>) toward nbCMs coated with different BCM preparations (20 min, 40 min, or 24 h) was evaluated by a transwell migration assay utilizing ThinCert<sup>®</sup> transwell PET membrane supports with 8 μm pore size. nbCM hydrated with RS was used as a control (Ctrl). (<b>a</b>,<b>c</b>,<b>e</b>) Representative images of fixed and stained cells that have migrated to the lower side of the filter in each of the experimental groups. Scale bar, 500 μm. (<b>b</b>,<b>d</b>,<b>f</b>) Quantification of the cell migration using the Image J software (version 1.50) measuring the area on the lower side of the filter covered with migrated cells. Data represent means ± SD from four independent experiments performed with (1) two independent BCM preparations, each used with two different cell donors for each of the two primary cell types, hBC and hPDLC, and (2) four independent BCM preparations used with the MG-63 cell line. Significant differences to the control, ** <span class="html-italic">p</span> &lt; 0.01, * <span class="html-italic">p</span> &lt; 0.05.</p>
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<p>Increased expression of adhesive marker genes in hBCs (<b>a</b>), hPDLCs (<b>b</b>), and MG-63 cells (<b>c</b>) grown on BCM-free nbCM (Ctrl) or nbCMs coated with either 20 min BCM, 40 min BCM, or 24 h BCM. Cells were cultured on the respective nbCM for 6 h followed by an extensive wash for complete removal of nonadherent cells from the membranes before total RNA was isolated and analyzed for the expression of adhesive marker genes (FN1, VCL, CD44, and ICAM1) by qRT-PCR. Values normalized to GAPDH are expressed relative to the values of control cells. Data represent means ± SD from four independent experiments performed with (1) two independent BCM preparations, each used with two different cell donors for each of the primary cell types, hBC and hPDLC, and (2) four independent BCM preparations used with the MG-63 cell line. Significant differences to the respective controls, *** <span class="html-italic">p</span> &lt; 0.001, ** <span class="html-italic">p</span> &lt; 0.01, * <span class="html-italic">p</span> &lt; 0.05. Morphological appearance and filamentous actin (F-actin) formation in hBCs, hPDLCs, and MG-63 cells, which, after initial attachment on control or BCM-coated nbCMs for 6 h, were detached and re-seeded on regular cell culture-treated plastic dishes for 24 h (<b>d</b>). Re-attached cells from all tested conditions were subjected to F-actin immunostaining using Alexa Fluor 488-labeled phalloidin (green). The cell nuclei were localized via DAPI co-stain (blue); a bright field (BF) image is also shown. Scale bar, 500 µm.</p>
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<p>(<b>a</b>–<b>c</b>) Proliferation rates of hBCs (<b>a</b>), hPDLCs (<b>b</b>), and MG-63 cells (<b>c</b>) grown on BCM-free nbCM (Ctrl) or nbCMs coated with either 20 min, 40 min, or 24 h BCM preparation were assessed by trypan blue dye-exclusion cell counting performed in a Countess™ II instrument on days 1, 3, 6, and 9 post-seeding. Data represent means ± SD from four independent experiments performed with (1) two independent BCM preparations, each used with two different cell donors for each of the primary cell types, hBC and hPDLC, and (2) four independent BCM preparations used with the MG-63 cell line. Significant differences to control cells at each individual time point unless otherwise indicated, *** <span class="html-italic">p</span> &lt; 0.001, ** <span class="html-italic">p</span> &lt; 0.01, * <span class="html-italic">p</span> &lt; 0.05. (<b>d</b>–<b>f</b>) Increased expression of proliferative marker genes in hBCs (<b>d</b>), hPDLCs (<b>e</b>), and MG-63 cells (<b>f</b>) grown on control or BCM-coated nbCMs. Cells were grown in the four tested conditions for 1 and 3 days before total RNA was isolated and analyzed for the expression of proliferative marker genes (MYBL2, BUB1, PLK1, and MKI67) by qRT-PCR. Values normalized to GAPDH are expressed relative to the values of control cells at day 1 (1d). Data represent means ± SD from four independent experiments performed with (1) two independent BCM preparations, each used with two different cell donors for each of the primary cell types, hBC and hPDLC, and (2) four independent BCM preparations used with the MG-63 cell line. Significant differences to the respective controls at day 1 unless otherwise indicated, *** <span class="html-italic">p</span> &lt; 0.001, ** <span class="html-italic">p</span> &lt; 0.01, * <span class="html-italic">p</span> &lt; 0.05.</p>
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<p>(<b>a</b>–<b>c</b>) Increased expression of early osteogenic marker genes in hBCs (<b>a</b>), hPDLCs (<b>b</b>), and MG-63 cells (<b>c</b>) grown on control or BCM-coated nbCMs. Cells were grown in the four tested conditions for 3 days before total RNA was isolated, purified, and analyzed for the expression of COL1A1, SPP1, RUNX2, and ALPL osteogenic markers by qRT-PCR. Values normalized to GAPDH are expressed relative to the values of control cells. Data represent means ± SD from four independent experiments performed with (1) two independent BCM preparations, each used with two different cell donors for each of the primary cell types, hBC and hPDLC, and (2) four independent BCM preparations used with the MG-63 cell line. Significant differences to the respective controls unless otherwise indicated, *** <span class="html-italic">p</span> &lt; 0.001, ** <span class="html-italic">p</span> &lt; 0.01, * <span class="html-italic">p</span> &lt; 0.05. (<b>d</b>) Increased alkaline phosphatase (ALP) activity in hBCs, hPDLCs, and MG-63 cells grown on control or BCM-coated nbCMs. Cells were grown as in (<b>a</b>–<b>c</b>) before ALP activity in the cell culture supernatants was measured by a fluorometric analysis based on the hydrolysis of 4-methylumbelliferyl phosphate by the ALP into the fluorescent product 4-methylumbelliferone. Data and statistical significance are expressed as in (<b>a</b>–<b>c</b>).</p>
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<p>Differential expression of intermediate and late osteogenic marker genes in osteoblast-like cell types grown on nbCM coated with short- versus long-term extracted BCM. hBCs (<b>a</b>), hPDLCs (<b>b</b>), and MG-63 cells (<b>c</b>) were grown on control or BCM-coated nbCMs for 3 days before total RNA was extracted, purified, and analyzed for the expression of DLX5, IBSP, BGLAP2, and PHEX osteogenic markers by qRT-PCR. Values normalized to GAPDH are expressed relative to the values of control cells. Data represent means ±SD from four independent experiments performed with (1) two independent BCM preparations, each used with two different cell donors for each of the primary cell types, hBC and hPDLC, and (2) four independent BCM preparations used with the MG-63 cell line. Significant differences to the respective controls unless otherwise indicated, *** <span class="html-italic">p</span> &lt; 0.001, ** <span class="html-italic">p</span> &lt; 0.01, * <span class="html-italic">p</span> &lt; 0.05.</p>
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24 pages, 1016 KiB  
Article
MILD: Minimizing Idle Listening Energy Consumption via Down-Clocking for Energy-Efficient Wi-Fi Communications
by Jae-Hyeon Park, Young-Joo Suh, Dongdeok Kim, Harim Lee, Hyeongtae Ahn and Young Deok Park
Sensors 2025, 25(4), 1155; https://doi.org/10.3390/s25041155 - 13 Feb 2025
Abstract
Mobile devices, such as smartphones and laptops, face energy consumption challenges due to battery limitations, with Wi-Fi being one of the major sources of energy consumption in these devices. The IEEE 802.11 standard addresses this issue with Power Saving Mode (PSM), which reduces [...] Read more.
Mobile devices, such as smartphones and laptops, face energy consumption challenges due to battery limitations, with Wi-Fi being one of the major sources of energy consumption in these devices. The IEEE 802.11 standard addresses this issue with Power Saving Mode (PSM), which reduces power consumption but increases latency. To mitigate this latency, Adaptive-PSM (A-PSM) dynamically switches between PSM and Constantly Awake Mode (CAM); however, the associated Idle Listening (IL) process still results in high energy consumption. Various strategies have been proposed to optimize IL time; however, Medium Access Control (MAC)-level contention and network delays limit their effectiveness. To overcome these limitations, we propose MILD (Minimizing Idle Listening energy consumption via Down-clocking), a novel scheme that reduces energy consumption without compromising throughput. MILD introduces specialized preambles for Packet Arrival Detection (PAD) and Device Address Recognition (DAR), allowing the client to operate in a down-clocked state during IL and switch to full clocking only when necessary. Experimental results demonstrate that MILD reduces energy consumption by up to 23.6% while maintaining a minimal throughput loss of 12.5%, outperforming existing schemes. Full article
(This article belongs to the Special Issue Energy-Efficient Communication Networks and Systems: 2nd Edition)
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<p>Example of down-clocking operation in a downstream scenario.</p>
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<p>Address overhead of down clocking.</p>
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<p>Normalized fraction of time spent in idle listening state across various traffic patterns and environments.</p>
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<p>Operation flow of MILD.</p>
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<p>Example illustrating the client’s operation in MILD.</p>
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<p>PAD accuracy results with USRP testbed.</p>
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<p>DAR false negative and false positive probability results with USRP testbed. (<b>a</b>) DAR false negative probability result with USRP testbed. (<b>b</b>) DAR false positive probability result with USRP testbed.</p>
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<p>Comparison of throughput loss of E-MiLi and MILD.</p>
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<p>Distribution of Tx, Rx, idle, and sleep states for smartphone devices under various traffic patterns.</p>
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<p>Comparison of energy consumption and efficiency improvement in various traffic patterns. (<b>a</b>) Comparison of energy consumption between A-PSM, E-MiLi, and MILD for various traffic patterns, (<b>b</b>) Comparison of energy efficiency improvement of MILD over E-MiLi by down-clocking factor.</p>
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21 pages, 5272 KiB  
Article
Study on the Optimization of an Extraction Process of Two Triterpenoid Saponins in the Root of Rosa laevigata Michx. and Their Protective Effect on Acute Lung Injury
by Jingya Mo, Qiaoyu Deng, Yuanyuan Huang, Xuegong Jia, Fengfeng Xie, Bei Zhou, Hongwei Gao, Yanchun Wu and Jingquan Yuan
Pharmaceuticals 2025, 18(2), 253; https://doi.org/10.3390/ph18020253 - 13 Feb 2025
Abstract
Objectives: Kajiichigoside F1 and rosamultin are natural triterpenoid saponins found in the root of Rosa laevigata Michx. These compounds are isomers, making their separation challenging. Nonetheless, they have been reported to exhibit significant anti-inflammatory activity, although their mechanism of action remains unclear. This [...] Read more.
Objectives: Kajiichigoside F1 and rosamultin are natural triterpenoid saponins found in the root of Rosa laevigata Michx. These compounds are isomers, making their separation challenging. Nonetheless, they have been reported to exhibit significant anti-inflammatory activity, although their mechanism of action remains unclear. This study aimed to optimize the extraction process of echinacoside and rosamultin from R. laevigata and to elucidate the anti-inflammatory mechanisms of these saponins in an LPS-induced acute lung injury (ALI) model. Methods: The extraction process was optimized using a single-factor experiment and the Box–Behnken response surface methodology, with the content of kajiichigoside F1, rosamultin, and their total content serving as evaluation indices. The acute lung injury model was induced by LPS, and lung tissue damage was assessed through hematoxylin and eosin (HE) staining. The secretion of relevant inflammatory factors was quantified using enzyme-linked immunosorbent assay (ELISA), and the expression levels of associated proteins were analyzed via Western blotting. Results: The optimal extraction conditions were determined to be an ethanol volume fraction of 80.0%, a solid–liquid ratio of 1:25, an extraction duration of 80 min, and three extraction cycles. Kajiichigoside F1 and rosamultin were found to mitigate alveolar inflammation in mice with acute lung injury (ALI) by effectively reducing the expression of the pro-inflammatory cytokines TNF-α and IL-6. Additionally, these compounds down-regulated the expression of phosphorylated NF-κB p65 and NF-κB IκBα proteins, thereby alleviating inflammatory symptoms. Conclusions: Kajiichigoside F1 and rosamultin attenuate the inflammatory response in acute lung injury induced by lipopolysaccharide (LPS) stimulation through modulation of the NF-κB signaling pathway. This study preliminarily elucidates their anti-inflammatory mechanism, suggesting that both compounds possess therapeutic potential for ALI. These findings provide significant guidance for the future development of active components derived from the root of R. laevigata and establish a foundation for enhancing the quality standards of its medicinal materials. Full article
(This article belongs to the Section Natural Products)
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Graphical abstract

Graphical abstract
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<p>Chemical structural formula of kajiichigoside F1 and rosamultin.</p>
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<p>Effects of various extraction techniques on the concentrations of kajiichigoside F1 and rosamultin in the root of <span class="html-italic">R. laevigata</span> (<span class="html-italic">n</span> = 3).</p>
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<p>Effects of varying ethanol concentrations on the levels of kajiichigoside F1 and rosamultin in the root of <span class="html-italic">R</span>. <span class="html-italic">laevigata</span> (<span class="html-italic">n</span> = 3).</p>
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<p>Effects of different solid–liquid ratios on the contents of kajiichigoside F1 and rosamultin in the root of <span class="html-italic">R. laevigata (n</span> = 3).</p>
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<p>Effects of different extraction time on the content of kajiichigoside F1 and rosamultin in the root of <span class="html-italic">R. laevigata</span> (<span class="html-italic">n</span> = 3).</p>
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<p>Effects of different extraction times on the content of kajiichigoside F1 and rosamultin in the root of <span class="html-italic">R. laevigata</span> (<span class="html-italic">n</span> = 3).</p>
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<p>The response surface diagram illustrates the influence of various factors on the extraction of kajiichigoside F1 and rosamultin from the root of <span class="html-italic">R. laevigata</span>. Utilizing Design-Expert 8.0.6 software, the response surface analysis was conducted to examine the interactions among these factors, and the corresponding response surface curves were generated. A steeper response surface indicates a more pronounced effect on the extraction rate. Specifically, the diagrams depict (<b>a</b>) the impact of ethanol volume fraction and solid–liquid ratio on the total content of kajiichigoside F1 and rosamultin; (<b>b</b>) the influence of extraction time and ethanol volume fraction; (<b>c</b>) the effects of the number of extraction times and ethanol volume fraction; (<b>d</b>) the interaction between extraction time and solid–liquid ratio; (<b>e</b>) the combined effects of the number of extraction times and solid–liquid ratio; and (<b>f</b>) the relationship between the number of extraction time and extraction times on the total content of kajiichigoside F1 and rosamultin.</p>
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<p>The response surface diagram illustrates the influence of various factors on the extraction of kajiichigoside F1 and rosamultin from the root of <span class="html-italic">R. laevigata</span>. Utilizing Design-Expert 8.0.6 software, the response surface analysis was conducted to examine the interactions among these factors, and the corresponding response surface curves were generated. A steeper response surface indicates a more pronounced effect on the extraction rate. Specifically, the diagrams depict (<b>a</b>) the impact of ethanol volume fraction and solid–liquid ratio on the total content of kajiichigoside F1 and rosamultin; (<b>b</b>) the influence of extraction time and ethanol volume fraction; (<b>c</b>) the effects of the number of extraction times and ethanol volume fraction; (<b>d</b>) the interaction between extraction time and solid–liquid ratio; (<b>e</b>) the combined effects of the number of extraction times and solid–liquid ratio; and (<b>f</b>) the relationship between the number of extraction time and extraction times on the total content of kajiichigoside F1 and rosamultin.</p>
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<p>Effects of kajiichigoside F1 and rosamultin on lung index in LPS-induced ALI mice. The lung index was analyzed by calculating the wet weight/body weight ratio of lung tissue in ALI mice. Results are expressed as mean ± standard deviation (S.D.). * means compared with the model group: ** <span class="html-italic">p</span> &lt; 0.01, *** <span class="html-italic">p</span> &lt; 0.001; # means compared with the normal group: # <span class="html-italic">p</span> &lt; 0.05, ### <span class="html-italic">p</span> &lt; 0.001.</p>
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<p>Effects of kajiichigoside F1 and rosamultin on the pathomorphology of lung tissue in LPS-induced ALI mice (HE staining, ×100). Representative images of lung tissue pathology were obtained by performing H&amp;E staining on each group of mice. Con: normal group; LPS: model group; Dex: dexamethasone group; kajiichigoside F1: low-, medium-, and high-dose group; rosamultin: low-, medium-, and high-dose group.</p>
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<p>The effect of kajiichigoside F1 and rosamultin on the inhibition of pro-inflammatory cytokine TNF-α in lung tissue of LPS-induced ALI mice. Results are expressed as mean ± standard deviation (S.D.). * means compared with the model group: * <span class="html-italic">p</span> &lt; 0.05, ** <span class="html-italic">p</span> &lt; 0.01, *** <span class="html-italic">p</span> &lt; 0.001; # means compared with the normal group: # <span class="html-italic">p</span> &lt; 0.05.</p>
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<p>The effect of kajiichigoside F1 and rosamultin on the inhibition of pro-inflammatory cytokine IL-6 in lung tissue of LPS-induced ALI mice. Results are expressed as mean ± standard deviation (S.D.). * means compared with the model group: ** <span class="html-italic">p</span> &lt; 0.01, *** <span class="html-italic">p</span> &lt; 0.001; # means compared with the normal group: # <span class="html-italic">p</span> &lt; 0.05, ## <span class="html-italic">p</span> &lt; 0.01, ### <span class="html-italic">p</span> &lt; 0.001.</p>
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<p>The inhibitory effect of kajiichigoside F1 on the expression of NF-κB pathway protein in LPS-induced ALI mice. Results are expressed as mean ± standard deviation (S.D.). * means compared with the model group: ** <span class="html-italic">p</span> &lt; 0.01, *** <span class="html-italic">p</span> &lt; 0.001; # means compared with the normal group: ## <span class="html-italic">p</span> &lt; 0.01, ### <span class="html-italic">p</span> &lt; 0.001.</p>
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<p>The inhibitory effect of rosamultin on the expression of NF-κB pathway proteins in LPS-induced ALI mice. Results are expressed as mean ± standard deviation (S.D.). * means compared with the model group: * <span class="html-italic">p</span> &lt; 0.05, ** <span class="html-italic">p</span> &lt; 0.01, *** <span class="html-italic">p</span> &lt; 0.001; # means compared with the normal group: ### <span class="html-italic">p</span> &lt; 0.001.</p>
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19 pages, 4765 KiB  
Article
Unraveling the Ancient Introgression History of Xanthoceras (Sapindaceae): Insights from Phylogenomic Analysis
by Jian He, Mingyang Li, Huanyu Wu, Jin Cheng and Lei Xie
Int. J. Mol. Sci. 2025, 26(4), 1581; https://doi.org/10.3390/ijms26041581 - 13 Feb 2025
Abstract
Ancient introgression is an infrequent evolutionary process often associated with conflicts between nuclear and organellar phylogenies. Determining whether such conflicts arise from introgression, incomplete lineage sorting (ILS), or other processes is essential to understanding plant diversification. Previous studies have reported phylogenetic discordance in [...] Read more.
Ancient introgression is an infrequent evolutionary process often associated with conflicts between nuclear and organellar phylogenies. Determining whether such conflicts arise from introgression, incomplete lineage sorting (ILS), or other processes is essential to understanding plant diversification. Previous studies have reported phylogenetic discordance in the placement of Xanthoceras, but its causes remain unclear. Here, we analyzed transcriptome data from 41 Sapindaceae samples to reconstruct phylogenies and investigate this discordance. While nuclear phylogenies consistently placed Xanthoceras as sister to subfam. Hippocastanoideae, plastid data positioned it as the earliest-diverging lineage within Sapindaceae. Our coalescent simulations suggest that this cyto-nuclear discordance is unlikely to be explained by ILS alone. HyDe and PhyloNet analyses provided strong evidence that Xanthoceras experienced ancient introgression, incorporating approximately 16% of its genetic material from ancestral subfam. Sapindoideae lineages. Morphological traits further support this evolutionary history, reflecting characteristics of both contributing subfamilies. Likely occurring during the Paleogene, this introgression represents a rare instance of cross-subfamily gene flow shaping the evolutionary trajectory of a major plant lineage. Our findings clarify the evolutionary history of Xanthoceras and underscore the role of ancient introgression in driving phylogenetic conflicts, offering a rare example of introgression-driven diversification in angiosperms. Full article
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Graphical abstract

Graphical abstract
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<p>Coalescent-based species tree inference from the Angiosperms353 and transcriptome nuclear data. The topology of the species tree was inferred using ASTRAL based on 138 nuclear genes. The numbers at the nodes denote local posterior probabilities (ASTRAL-lpp), with unmarked nodes indicating a probability of 1.00. The colors of the tree branches correspond to the four subfamilies of Sapindaceae: red for subfam. Dodonaeoideae, green for subfam. Xanthoceroideae, brown for subfam. Hippocastanoideae, and blue for subfam. Sapindoideae. Support values for the branches are indicated by colored circles, where red circles denote nodes with lower support (ASTRAL-lpp &lt; 60%) and blue circles denote nodes with higher support (ASTRAL-lpp ≥ 60%). The sample names are color-coded to indicate data sources, with black labels representing species analyzed using the Angiosperms353 dataset from Buerki et al. [<a href="#B30-ijms-26-01581" class="html-bibr">30</a>] and green labels representing species sampled from this study’s transcriptome dataset.</p>
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<p>A chronogram of sapindaceae inferred from single-copy orthologous genes using the full-coalescent approach. Blue numbers on the nodes represent the median estimated divergence times in million years ago (Mya), while red numbers indicate the posterior probability support for the topology in the Bayesian inference (Using StarBeast3). Transparent lines over the nodes represent the 95% highest posterior density (HPD) intervals for the divergence times.</p>
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<p>Comparison of the Sapindaceae plastid and nuclear phylogenies. On the left, the Sapindaceae phylogeny, inferred from complete plastid genome sequences using maximum likelihood (ML) with the GTR+G model and Bayesian inference (BI) using MrBayes, is shown. Branches with ML bootstrap percentages less than 100% or BI posterior probability values less than 1 are indicated with the corresponding values. Red numbers in brackets represent the likelihood values indicating the probability of the plastid genome phylogeny topology under scenarios without interspecific gene flow events (considering only incomplete lineage sorting) based on the multispecies coalescent model simulated in Phybase. The ML phylogram is displayed below on the left. On the right, the full coalescence-based species tree topology, inferred using StarBeast3, is presented. Branches with posterior probabilities less than 1.0 are indicated with numbers. The coalescent chronogram is displayed below on the right.</p>
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<p>Comprehensive full coalescence-based species tree topology illustrating gene discordance. The species tree is depicted with heavy black lines. The gray-colored trees (cloud-tree) were sampled from 86 single-copy orthologous nuclear genes (excluding those with missing taxa) and constructed using RAxML. Pie charts at each node display the proportions of tree topologies that are concordant and discordant with the overarching species tree, providing a visual quantification of phylogenetic agreement and conflict.</p>
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<p>Interspecific gene flow detection based on HyDe (<b>A</b>) and PhyloNet (<b>B</b>) results. (<b>A</b>) On the left, the detected clade (<span class="html-italic">Xanthoceras</span>) is marked with a red star on the cladogram. The small blue squares in the heatmap indicate the corresponding interspecific gene flow signals detected by HyDe. The intensity of the blue color represents the inheritance probabilities of the corresponding taxa on the left axis. When small squares of the same color intensity form a larger square in the heatmap, and their corresponding taxa on the coordinate axis form a monophyletic clade, it suggests that the most recent common ancestor of the clade may be one of the parents. On the right, a schematic diagram of a phylogenetic network is shown based on the left cladogram and heatmap. Numerical values indicate inheritance probabilities. (<b>B</b>) Interspecific gene flow within Sapindaceae clades as estimated by PhyloNet. The results show maximum pseudolikelihood trees (right), with one best-allowed reticulation based on the pseudo-log-likelihood scores (left).</p>
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