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19 pages, 1672 KiB  
Article
Hibiscus syriacus L. Exhibits Cardioprotective Activity via Anti-Inflammatory and Antioxidant Mechanisms in an In Vitro Model of Heart Failure
by Hung-Hsin Chao, Tzu-Hurng Cheng, Chun-Chao Chen, Ju-Chi Liu, Jin-Jer Chen and Li-Chin Sung
Life 2025, 15(8), 1229; https://doi.org/10.3390/life15081229 (registering DOI) - 3 Aug 2025
Abstract
Hibiscus syriacus L. (HS), native to Eastern and Southern Asia, has been traditionally used in Asian herbal medicine for its anticancer, antimicrobial, and anti-inflammatory properties. Despite these recognized bioactivities, its potential cardioprotective effects, particularly in the setting of heart failure (HF), remain largely [...] Read more.
Hibiscus syriacus L. (HS), native to Eastern and Southern Asia, has been traditionally used in Asian herbal medicine for its anticancer, antimicrobial, and anti-inflammatory properties. Despite these recognized bioactivities, its potential cardioprotective effects, particularly in the setting of heart failure (HF), remain largely unexplored. This study aimed to investigate the effects of HS extracts and its bioactive constituents on angiotensin II (Ang II)-induced cardiac injury using an in vitro model with H9c2 rat cardiomyocytes. Cells exposed to Ang II were pretreated with HS extracts, and assays were performed to assess cell viability, reactive oxygen species (ROS) generation, protein synthesis, and secretion of inflammatory mediators, including tumor necrosis factor-alpha, interleukin 1β (IL-1β), and interleukin 6 (IL-6), as well as chemokine (CCL20) and HF-related biomarkers, such as brain natriuretic peptide (BNP) and endothelin-1. The results demonstrated that HS extracts significantly and dose-dependently attenuated Ang II-induced ROS accumulation and suppressed the secretion of pro-inflammatory cytokines, chemokines, BNP, and endothelin-1. Additionally, HS and its purified components inhibited Ang II-induced protein synthesis, indicating anti-hypertrophic effects. Collectively, these findings highlight the antioxidative, anti-inflammatory, and antihypertrophic properties of HS in the context of Ang II-induced cardiac injury, suggesting that HS may represent a promising adjunctive therapeutic candidate for HF management. Further in vivo studies and mechanistic investigations are warranted to validate its clinical potential. Full article
(This article belongs to the Special Issue Advancements in Heart Failure Research: 2nd Edition)
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11 pages, 876 KiB  
Article
Body Composition Changes in Hospitalized Patients with Community-Acquired Pneumonia
by Ryuji Sugiya, Osamu Nishiyama, Masashi Shiraishi, Kazuya Yoshikawa, Kyuya Gose, Ryo Yamazaki, Takashi Oomori, Akiko Sano, Shinichi Arizono, Yasushi Uchiyama, Yuji Higashimoto and Hisako Matsumoto
J. Clin. Med. 2025, 14(15), 5460; https://doi.org/10.3390/jcm14155460 (registering DOI) - 3 Aug 2025
Abstract
Background: The influence of hospitalization owing to pneumonia on changes in body composition has not been specifically reported. We conducted a prospective cohort study of patients with community-acquired pneumonia (CAP) requiring hospitalization to test the hypothesis that hospitalization affects body composition. Methods [...] Read more.
Background: The influence of hospitalization owing to pneumonia on changes in body composition has not been specifically reported. We conducted a prospective cohort study of patients with community-acquired pneumonia (CAP) requiring hospitalization to test the hypothesis that hospitalization affects body composition. Methods: Sixty-four consecutive patients with CAP were recruited. Body composition was measured within 24 h of admission and 24 h before discharge using bioelectrical impedance analysis. The association between changes in body composition and variables obtained at admission was investigated. Index values were calculated as weight divided by height squared. Results: The mean age of the patients was 76.0 ± 8.7 years (78.1% males). The median length of hospitalization was 12.0 days. Weight, body mass index (BMI), skeletal muscle (SM), SM index, fat-free mass (FFM), and FFM index significantly decreased (p < 0.001 for each), but fat mass (FM) and FM index did not. The serum total protein level was the only independent predictor of the lowest quartile of change in SM index (<−0.4) after adjusting for age and sex (p = 0.004). Conclusions: In summary, weight and BMI significantly decreased during hospitalization in patients with CAP, which was attributed to SM reduction. Patients with low serum total protein levels on admission were at risk of an accelerated decrease in the SM index. Nutritional intervention and rehabilitation are important for these patients. Full article
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16 pages, 577 KiB  
Systematic Review
Blood Biomarkers as Optimization Tools for Computed Tomography in Mild Traumatic Brain Injury Management in Emergency Departments: A Systematic Review
by Ángela Caballero Ballesteros, María Isabel Alonso Gallardo and Juan Mora-Delgado
J. Pers. Med. 2025, 15(8), 350; https://doi.org/10.3390/jpm15080350 (registering DOI) - 3 Aug 2025
Abstract
Background/Objectives: Traumatic brain injury (TBI), especially mild TBI (mTBI), is frequently caused by traffic accidents, falls, or sports injuries. Although computed tomography (CT) is the gold standard for diagnosis, overuse can lead to unnecessary radiation exposure, increased healthcare costs, and emergency department [...] Read more.
Background/Objectives: Traumatic brain injury (TBI), especially mild TBI (mTBI), is frequently caused by traffic accidents, falls, or sports injuries. Although computed tomography (CT) is the gold standard for diagnosis, overuse can lead to unnecessary radiation exposure, increased healthcare costs, and emergency department saturation. Blood-based biomarkers have emerged as potential tools to optimize CT scan use. This systematic review aims to evaluate recent evidence on the role of specific blood biomarkers in guiding CT decisions in patients with mTBI. Methods: A systematic search was conducted in the PubMed, Cochrane, and CINAHL databases for studies published between 2020 and 2024. Inclusion criteria focused on adult patients with mTBI evaluated using both CT imaging and at least one of the following biomarkers: glial fibrillary acidic protein (GFAP), ubiquitin carboxy-terminal hydrolase L1 (UCH-L1), and S100 calcium-binding protein B (S100B). After screening, six studies were included in the final review. Results: All included studies reported high sensitivity and negative predictive value for the selected biomarkers in detecting clinically relevant intracranial lesions. GFAP and UCH-L1, particularly in combination, consistently identified low-risk patients who could potentially forgo CT scans. While S100B also showed high sensitivity, discrepancies in cutoff values across studies highlighted the need for harmonization. Conclusions: Blood biomarkers such as GFAP, UCH-L1, and S100B demonstrate strong potential to reduce unnecessary CT imaging in mTBI by identifying patients at low risk of significant brain injury. Future research should focus on standardizing biomarker thresholds and validating protocols to support their integration into clinical practice guidelines. Full article
21 pages, 2818 KiB  
Review
Structuring the Future of Cultured Meat: Hybrid Gel-Based Scaffolds for Edibility and Functionality
by Sun Mi Mi Zo, Ankur Sood, So Yeon Yeon Won, Soon Mo Mo Choi and Sung Soo Soo Han
Gels 2025, 11(8), 610; https://doi.org/10.3390/gels11080610 (registering DOI) - 3 Aug 2025
Abstract
Cultured meat is emerging as a sustainable alternative to conventional animal agriculture, with scaffolds playing a central role in supporting cellular attachment, growth, and tissue maturation. This review focuses on the development of gel-based hybrid biomaterials that meet the dual requirements of biocompatibility [...] Read more.
Cultured meat is emerging as a sustainable alternative to conventional animal agriculture, with scaffolds playing a central role in supporting cellular attachment, growth, and tissue maturation. This review focuses on the development of gel-based hybrid biomaterials that meet the dual requirements of biocompatibility and food safety. We explore recent advances in the use of naturally derived gel-forming polymers such as gelatin, chitosan, cellulose, alginate, and plant-based proteins as the structural backbone for edible scaffolds. Particular attention is given to the integration of food-grade functional additives into hydrogel-based scaffolds. These include nanocellulose, dietary fibers, modified starches, polyphenols, and enzymatic crosslinkers such as transglutaminase, which enhance mechanical stability, rheological properties, and cell-guidance capabilities. Rather than focusing on fabrication methods or individual case studies, this review emphasizes the material-centric design strategies for building scalable, printable, and digestible gel scaffolds suitable for cultured meat production. By systemically evaluating the role of each component in structural reinforcement and biological interaction, this work provides a comprehensive frame work for designing next-generation edible scaffold systems. Nonetheless, the field continues to face challenges, including structural optimization, regulatory validation, and scale-up, which are critical for future implementation. Ultimately, hybrid gel-based scaffolds are positioned as a foundational technology for advancing the functionality, manufacturability, and consumer readiness of cultured meat products, distinguishing this work from previous reviews. Unlike previous reviews that have focused primarily on fabrication techniques or tissue engineering applications, this review provides a uniquely food-centric perspective by systematically evaluating the compositional design of hybrid hydrogel-based scaffolds with edibility, scalability, and consumer acceptance in mind. Through a comparative analysis of food-safe additives and naturally derived biopolymers, this review establishes a framework that bridges biomaterials science and food engineering to advance the practical realization of cultured meat products. Full article
(This article belongs to the Special Issue Food Hydrocolloids and Hydrogels: Rheology and Texture Analysis)
17 pages, 1164 KiB  
Systematic Review
Effects of Pomegranate Juice on Androgen Levels, Inflammation and Lipid Profile in Polycystic Ovary Syndrome: A Systematic Review and Meta-Analysis
by Vitória Silveira, Pamela Braz, Antonio Jose Grande, Tamy Colonetti, Maria Laura Rodrigues Uggioni, Gabriele da Silveira Prestes, Leonardo Roever, Valdemira Santina Dagostin and Maria Inês da Rosa
J. Clin. Med. 2025, 14(15), 5458; https://doi.org/10.3390/jcm14155458 (registering DOI) - 3 Aug 2025
Abstract
Background/Objectives: Polycystic ovary syndrome (PCOS) is a multifactorial endocrine disorder frequently associated with metabolic and inflammatory disturbances. Due to its antioxidant and anti-inflammatory properties, pomegranate juice has been proposed as a potential adjunctive therapy in managing PCOS. To evaluate the effects of pomegranate [...] Read more.
Background/Objectives: Polycystic ovary syndrome (PCOS) is a multifactorial endocrine disorder frequently associated with metabolic and inflammatory disturbances. Due to its antioxidant and anti-inflammatory properties, pomegranate juice has been proposed as a potential adjunctive therapy in managing PCOS. To evaluate the effects of pomegranate juice on hormonal, inflammatory, and lipid parameters and body mass index (BMI) in women with PCOS. Methods: A systematic review and meta-analysis of randomized controlled trials (RCTs) was conducted following PRISMA guidelines. Comprehensive searches were performed in electronic databases including Medline, Scopus, Web of Science, Cochrane CENTRAL, and Embase from inception to July 2025, using keywords and MeSH terms related to “polycystic ovary syndrome” and “pomegranate juice” without language restrictions. The primary outcomes were changes in serum testosterone, luteinizing hormone (LH), high-sensitivity C-reactive protein (hs-CRP), lipid profile parameters (HDL, LDL, triglycerides, and total cholesterol), and body mass index (BMI). Results: Four RCTs published between 2020 and 2023, encompassing 128 women with PCOS, were included. The meta-analysis revealed significant reductions in testosterone (MD: −0.05; 95% CI: −0.07 to −0.03; p < 0.0001; I2 = 0%, two studies, 85 participants) and hs-CRP (SMD: −0.85; 95% CI: −1.35 to −0.35; p = 0.0009; I2 = 20%, two studies, 85 participants), along with increases in HDL (MD: 6.21; 95% CI: 2.43 to 10.00; p = 0.001; I2 = 0%, two studies, 85 participants) and reductions in triglycerides (MD: −23.30; 95% CI: −45.19 to −1.42; p = 0.04; I2 = 0%, two studies, 85 participants). No significant changes were observed in LH, LDL, total cholesterol, or BMI. Conclusions: Pomegranate juice demonstrates promising effects as an adjunctive intervention in women with PCOS, improving androgen levels, inflammatory markers, and certain lipid parameters. Further long-term studies are needed to confirm these findings. Full article
(This article belongs to the Section Clinical Laboratory Medicine)
21 pages, 3431 KiB  
Article
Synthesis and Antibacterial Evaluation of an Indole Triazole Conjugate with In Silico Evidence of Allosteric Binding to Penicillin-Binding Protein 2a
by Vidyasrilekha Sanapalli, Bharat Kumar Reddy Sanapalli and Afzal Azam Mohammed
Pharmaceutics 2025, 17(8), 1013; https://doi.org/10.3390/pharmaceutics17081013 (registering DOI) - 3 Aug 2025
Abstract
Background: Antibacterial resistance (ABR) poses a major challenge to global health, with methicillin-resistant Staphylococcus aureus (MRSA) being one of the prominent multidrug-resistant strains. MRSA has developed resistance through the expression of Penicillin-Binding Protein 2a (PBP2a), a key transpeptidase enzyme involved in bacterial [...] Read more.
Background: Antibacterial resistance (ABR) poses a major challenge to global health, with methicillin-resistant Staphylococcus aureus (MRSA) being one of the prominent multidrug-resistant strains. MRSA has developed resistance through the expression of Penicillin-Binding Protein 2a (PBP2a), a key transpeptidase enzyme involved in bacterial cell wall biosynthesis. Objectives: The objective was to design and characterize a novel small-molecule inhibitor targeting PBP2a as a strategy to combat MRSA. Methods: We synthesized a new indole triazole conjugate (ITC) using eco-friendly and click chemistry approaches. In vitro antibacterial tests were performed against a panel of strains to evaluate the ITC antibacterial potential. Further, a series of in silico evaluations like molecular docking, MD simulations, free energy landscape (FEL), and principal component analysis (PCA) using the crystal structure of PBP2a (PDB ID: 4CJN), in order to predict the mechanism of action, binding mode, structural stability, and energetic profile of the 4CJN-ITC complex. Results: The compound ITC exhibited noteworthy antibacterial activity, which effectively inhibited the selected strains. Binding score and energy calculations demonstrated high affinity of ITC for the allosteric site of PBP2a and significant interactions responsible for complex stability during MD simulations. Further, FEL and PCA provided insights into the conformational behavior of ITC. These results gave the structural clues for the inhibitory action of ITC on the PBP2a. Conclusions: The integrated in vitro and in silico studies corroborate the potential of ITC as a promising developmental lead targeting PBP2a in MRSA. This study demonstrates the potential usage of rational drug design approaches in addressing therapeutic needs related to ABR. Full article
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22 pages, 11874 KiB  
Article
Bactericidal Activities of Nanoemulsion Containing Piper betle L. Leaf and Hydroxychavicol Against Avian Pathogenic Escherichia coli and Modelling Simulation of Hydroxychavicol Against Bacterial Cell Division Proteins
by Kunchaphorn Ratchasong, Phirabhat Saengsawang, Gorawit Yusakul, Fonthip Makkliang, Hemanth Kumar Lakhanapuram, Phitchayapak Wintachai, Thotsapol Thomrongsuwannakij, Ozioma Forstinus Nwabor, Veerasak Punyapornwithaya, Chonticha Romyasamit and Watcharapong Mitsuwan
Antibiotics 2025, 14(8), 788; https://doi.org/10.3390/antibiotics14080788 (registering DOI) - 3 Aug 2025
Abstract
Background: Avian pathogenic Escherichia coli (APEC) is a leading cause of colibacillosis in poultry. Piper betle L. is a medicinal plant rich in bioactive compounds including hydroxychavicol that possess potent antibacterial activity. This study aimed to investigate the efficacy of a P. [...] Read more.
Background: Avian pathogenic Escherichia coli (APEC) is a leading cause of colibacillosis in poultry. Piper betle L. is a medicinal plant rich in bioactive compounds including hydroxychavicol that possess potent antibacterial activity. This study aimed to investigate the efficacy of a P. betle L. leaf nanoemulsion (NEPE) and hydroxychavicol against multidrug-resistant APEC isolates. Methods: In vitro and in silico analysis of NEPE and hydroxychavicol against APEC were determined. Results: The nanoemulsion exhibited potent antibacterial activity, with MIC and MBC values of 0.06–0.25% v/v and 0.125–0.25% v/v, respectively. The MIC and MBC values of hydroxychavicol against isolates ranged from 0.25 to 1.0 mg/mL. A time–kill assays revealed rapid bactericidal effects of both compounds, achieving a ≥3-log reduction within 4 h at 4 × MIC. Scanning electron microscopy demonstrated that APEC cells treated with hydroxychavicol exhibited filamentous cells with incomplete septa. Molecular docking and dynamics simulations of hydroxychavicol against APEC cell division proteins were investigated. According to the binding energy, hydroxychavicol exhibited the highest affinity with ZapE, FtsW, FtsX, FtsZ, and FtsA, respectively. However, the FtsA protein showed the least protein conformational change throughout the 5000 ns simulation, reflecting a highly stable conformation. Conclusions: These confirm the potential stability of protein and ligand, as supported by molecular dynamics simulation. The results suggested the potential of NEPE and hydroxychavicol, which may have promising antibacterial potential that can be used to inhibit APEC growth. Full article
(This article belongs to the Special Issue Antimicrobial Extracts and Compounds Derived from Plants)
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13 pages, 6042 KiB  
Article
Whey Protein–Quercetin–Gellan Gum Complexes Prepared Using pH-Shift Treatment: Structural and Functional Properties
by Na Guo, Xin Zhou, Ganghua Zhou, Yimeng Zhang, Guoqing Yu, Yangliu Liu, Beibei Li, Fangyan Zhang and Guilan Zhu
Foods 2025, 14(15), 2720; https://doi.org/10.3390/foods14152720 (registering DOI) - 3 Aug 2025
Abstract
The objectives of this study were to prepare whey protein–quercetin–gellan gum conjugates using the pH-shift method and to evaluate the impacts of varying pH values and quercetin concentrations on the interaction mechanisms and functional characteristics of the complexes. Spectroscopic analyses (fluorescence, UV-vis, and [...] Read more.
The objectives of this study were to prepare whey protein–quercetin–gellan gum conjugates using the pH-shift method and to evaluate the impacts of varying pH values and quercetin concentrations on the interaction mechanisms and functional characteristics of the complexes. Spectroscopic analyses (fluorescence, UV-vis, and FT-IR) revealed that new complexes formed under alkaline conditions. Notably, an increasing quercetin concentration led to a reduction in complex particle size and an increase in the zeta potential value, with these effects being more pronounced under alkaline conditions. The particle size was 425.7 nm, and the zeta potential value was −30.00 mV at a quercetin addition concentration of 15 umol/g protein. Additionally, the complexes formed under alkaline conditions exhibited superior foaming capacity, emulsification properties, and significantly enhanced free radical scavenging activity. The complex’s DPPH and ABTS radical scavenging rates rose by 41.57% and 57.69%, respectively. This study provides theoretical foundations and practical insights for developing protein—polyphenol systems, offering significant implications for the application of quercetin functional foods and supplements in the food science and pharmaceutical industries. Full article
(This article belongs to the Special Issue Oil and Protein Engineering and Its Applications in Food Industry)
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10 pages, 216 KiB  
Perspective
Silicon Is the Next Frontier in Plant Synthetic Biology
by Aniruddha Acharya, Kaitlin Hopkins and Tatum Simms
SynBio 2025, 3(3), 12; https://doi.org/10.3390/synbio3030012 (registering DOI) - 3 Aug 2025
Abstract
Silicon has a striking similarity to carbon and is found in plant cells. However, there is no specific role that has been assigned to silicon in the life cycle of plants. The amount of silicon in plant cells is species specific and can [...] Read more.
Silicon has a striking similarity to carbon and is found in plant cells. However, there is no specific role that has been assigned to silicon in the life cycle of plants. The amount of silicon in plant cells is species specific and can reach levels comparable to macronutrients. Silicon is used extensively in artificial intelligence, nanotechnology, and the digital revolution, and thus can serve as an informational molecule such as nucleic acids. The diverse potential of silicon to bond with different chemical species is analogous to carbon; thus, it can serve as a structural candidate similar to proteins. The discovery of large amounts of silicon on Mars and the moon, along with the recent development of enzyme that can incorporate silicon into organic molecules, has propelled the theory of creating silicon-based life. The bacterial cytochrome has been modified through directed evolution such that it could cleave silicon–carbon bonds in organo-silicon compounds. This consolidates the idea of utilizing silicon in biomolecules. In this article, the potential of silicon-based life forms has been hypothesized, along with the reasoning that autotrophic virus-like particles could be used to investigate such potential. Such investigations in the field of synthetic biology and astrobiology will have corollary benefits for Earth in the areas of medicine, sustainable agriculture, and environmental sustainability. Full article
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15 pages, 1407 KiB  
Article
Expression of Recombinant Hirudin in Bacteria and Yeast: A Comparative Approach
by Zhongjie Wang, Dominique Böttcher, Uwe T. Bornscheuer and Christian Müller
Methods Protoc. 2025, 8(4), 89; https://doi.org/10.3390/mps8040089 (registering DOI) - 3 Aug 2025
Abstract
The expression of recombinant proteins in heterologous hosts is a common strategy to obtain larger quantities of the “protein of interest” (POI) for scientific, therapeutic or commercial purposes. However, the experimental success of such an approach critically depends on the choice of an [...] Read more.
The expression of recombinant proteins in heterologous hosts is a common strategy to obtain larger quantities of the “protein of interest” (POI) for scientific, therapeutic or commercial purposes. However, the experimental success of such an approach critically depends on the choice of an appropriate host system to obtain biologically active forms of the POI. The correct folding of the molecule, mediated by disulfide bond formation, is one of the most critical steps in that process. Here we describe the recombinant expression of hirudin, a leech-derived anticoagulant and thrombin inhibitor, in the yeast Komagataella phaffii (formerly known and mentioned throughout this publication as Pichia pastoris) and in two different strains of Escherichia coli, one of them being especially designed for improved disulfide bond formation through expression of a protein disulfide isomerase. Cultivation of the heterologous hosts and expression of hirudin were performed at different temperatures, ranging from 22 to 42 °C for the bacterial strains and from 20 to 30 °C for the yeast strain, respectively. The thrombin-inhibitory potencies of all hirudin preparations were determined using the thrombin time coagulation assay. To our surprise, the hirudin preparations of P. pastoris were considerably less potent as thrombin inhibitors than the respective preparations of both E. coli strains, indicating that a eukaryotic background is not per se a better choice for the expression of a biologically active eukaryotic protein. The hirudin preparations of both E. coli strains exhibited comparable high thrombin-inhibitory potencies when the strains were cultivated at their respective optimal temperatures, whereas lower or higher cultivation temperatures reduced the inhibitory potencies. Full article
(This article belongs to the Section Molecular and Cellular Biology)
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16 pages, 1961 KiB  
Article
A Novel Glycosylated Ferulic Acid Conjugate: Synthesis, Antioxidative Neuroprotection Activities In Vitro, and Alleviation of Cerebral Ischemia–Reperfusion Injury (CIRI) In Vivo
by Jian Chen, Yongjun Yuan, Litao Tong, Manyou Yu, Yongqing Zhu, Qingqing Liu, Junling Deng, Fengzhang Wang, Zhuoya Xiang and Chen Xia
Antioxidants 2025, 14(8), 953; https://doi.org/10.3390/antiox14080953 (registering DOI) - 3 Aug 2025
Abstract
Antioxidative neuroprotection is effective at preventing ischemic stroke (IS). Ferulic acid (FA) offers benefits in the treatment of many diseases, mostly due to its antioxidant activities. In this study, a glycosylated ferulic acid conjugate (FA-Glu), with 1,2,3-triazole as a linker and bioisostere between [...] Read more.
Antioxidative neuroprotection is effective at preventing ischemic stroke (IS). Ferulic acid (FA) offers benefits in the treatment of many diseases, mostly due to its antioxidant activities. In this study, a glycosylated ferulic acid conjugate (FA-Glu), with 1,2,3-triazole as a linker and bioisostere between glucose at the C6 position and FA at the C4 position, was designed and synthesized. The hydrophilicity and chemical stability of FA-Glu were tested. FA-Glu’s protection against DNA oxidative cleavage was tested using pBR322 plasmid DNA under the Fenton reaction. The cytotoxicity of FA-Glu was examined via the PC12 cell and bEnd.3 cell tests. Antioxidative neuroprotection was evaluated, in vitro, via a H2O2-induced PC12 cell test, measuring cell viability and ROS levels. Antioxidative alleviation of cerebral ischemia–reperfusion injury (CIRI), in vivo, was evaluated using a rat middle cerebral artery occlusion (MCAO) model. The results indicated that FA-Glu was water-soluble (LogP −1.16 ± 0.01) and chemically stable. FA-Glu prevented pBR322 plasmid DNA cleavage induced via •OH radicals (SC% 88.00%). It was a non-toxic agent based on PC12 cell and bEnd.3 cell tests results. FA-Glu significantly protected against H2O2-induced oxidative damage in the PC12 cell (cell viability 88.12%, 100 μM) and inhibited excessive cell ROS generation (45.67% at 100 μM). FA-Glu significantly reduced the infarcted brain areas measured using TTC stain observation, quantification (FA-Glu 21.79%, FA 28.49%, I/R model 43.42%), and H&E stain histological observation. It sharply reduced the MDA level (3.26 nmol/mg protein) and significantly increased the GSH level (139.6 nmol/mg protein) and SOD level (265.19 U/mg protein). With superior performance to FA, FA-Glu is a safe agent with effective antioxidative DNA and neuronal protective actions and an ability to alleviate CIRI, which should help in the prevention of IS. Full article
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12 pages, 3410 KiB  
Article
Nasal and Ocular Immunization with Bacteriophage Virus-Like Particle Vaccines Elicits Distinct Systemic and Mucosal Antibody Profiles
by Andzoa N. Jamus, Zoe E. R. Wilton, Samantha D. Armijo, Julian Flanagan, Isabella G. Romano, Susan B. Core and Kathryn M. Frietze
Vaccines 2025, 13(8), 829; https://doi.org/10.3390/vaccines13080829 (registering DOI) - 3 Aug 2025
Abstract
Background/Objectives: Intramuscular immunization elicits systemic IgG and is the primary route of vaccine administration in humans. However, there is growing interest in utilizing other routes of administration to tailor antibody profiles, increase immunity at primary sites of infection, simplify administration, and eliminate [...] Read more.
Background/Objectives: Intramuscular immunization elicits systemic IgG and is the primary route of vaccine administration in humans. However, there is growing interest in utilizing other routes of administration to tailor antibody profiles, increase immunity at primary sites of infection, simplify administration, and eliminate needle waste. Here, we investigated the antibody profiles elicited by immunization with bacteriophage virus-like particle vaccine platforms at various routes of administration. Methods: We chose two model bacteriophage vaccines for investigation: bacteriophage MS2 virus-like particles (VLPs) recombinantly displaying a short, conserved peptide from Chlamydia trachomatis major outer membrane protein (MS2) and bacteriophage Qβ VLPs displaying oxycodone through chemical conjugation (Qβ). We comprehensively characterized the antibodies elicited systemically and at various mucosal sites when the vaccines were administered intramuscularly, intranasally or periocularly with or without an intramuscular prime using various prime/boost schemes. Results: Intranasal and periocular immunization elicited robust mucosal and systemic IgA responses for both MS2 and Qβ. The intramuscular prime followed by intranasal or periocular boosts elicited broad antibody responses, and increased antibodies titers at certain anatomical sites. Conclusions: These findings demonstrate the tractability of bacteriophage VLP-based vaccines in generating specific antibody profiles based on the prime–boost regimen and route of administration. Full article
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18 pages, 4468 KiB  
Article
Proteomic and Functional Analysis Reveals Temperature-Driven Immune Evasion Strategies of Streptococcus iniae in Yellowfin Seabream (Acanthopagrus latus)
by Yanjian Yang, Guanrong Zhang, Ruilong Xu, Yiyang Deng, Zequan Mo, Yanwei Li and Xueming Dan
Biology 2025, 14(8), 986; https://doi.org/10.3390/biology14080986 (registering DOI) - 2 Aug 2025
Abstract
Streptococcus iniae (S. iniae) is a globally significant aquatic pathogen responsible for severe economic losses in aquaculture. While the S. iniae infection often exhibits distinct seasonal patterns strongly correlated with water temperature, there is limited knowledge regarding the temperature-dependent immune evasion [...] Read more.
Streptococcus iniae (S. iniae) is a globally significant aquatic pathogen responsible for severe economic losses in aquaculture. While the S. iniae infection often exhibits distinct seasonal patterns strongly correlated with water temperature, there is limited knowledge regarding the temperature-dependent immune evasion strategies of S. iniae. Our results demonstrated a striking temperature-dependent virulence phenotype, with significantly higher A. latus mortality rates observed at high temperature (HT, 33 °C) compared to low temperature (LT, 23 °C). Proteomic analysis revealed temperature-dependent upregulation of key virulence factors, including streptolysin S-related proteins (SagG, SagH), antioxidant-related proteins (SodA), and multiple capsular polysaccharide (cps) synthesis proteins (cpsD, cpsH, cpsL, cpsY). Flow cytometry analysis showed that HT infection significantly reduced the percentage of lymphocyte and myeloid cell populations in the head kidney leukocytes of A. latus, which was associated with elevated caspase-3/7 expression and increased apoptosis. In addition, HT infection significantly inhibited the release of reactive oxygen species (ROS) but not nitric oxide (NO) production. Using S. iniae cps-deficient mutant, Δcps, we demonstrated that the cps is essential for temperature-dependent phagocytosis resistance in S. iniae, as phagocytic activity against Δcps remained unchanged across temperatures, while NS-1 showed significantly reduced uptake at HT. These findings provide new insights into the immune evasion of S. iniae under thermal regulation, deepening our understanding of the thermal adaptation of aquatic bacterial pathogens. Full article
(This article belongs to the Special Issue Aquatic Economic Animal Breeding and Healthy Farming)
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16 pages, 2227 KiB  
Article
Physiological and Transcriptomic Mechanisms Underlying Vitamin C-Mediated Cold Stress Tolerance in Grafted Cucumber
by Panpan Yu, Junkai Wang, Xuyang Zhang, Zhenglong Weng, Kaisen Huo, Qiuxia Yi, Chenxi Wu, Sunjeet Kumar, Hao Gao, Lin Fu, Yanli Chen and Guopeng Zhu
Plants 2025, 14(15), 2398; https://doi.org/10.3390/plants14152398 (registering DOI) - 2 Aug 2025
Abstract
Cucumbers (Cucumis sativus L.) are highly sensitive to cold, but grafting onto cold-tolerant rootstocks can enhance their low-temperature resilience. This study investigates the physiological and molecular mechanisms by which exogenous vitamin C (Vc) mitigates cold stress in grafted cucumber seedlings. Using cucumber [...] Read more.
Cucumbers (Cucumis sativus L.) are highly sensitive to cold, but grafting onto cold-tolerant rootstocks can enhance their low-temperature resilience. This study investigates the physiological and molecular mechanisms by which exogenous vitamin C (Vc) mitigates cold stress in grafted cucumber seedlings. Using cucumber ‘Chiyu 505’ as the scion and pumpkin ‘Chuangfan No.1’ as the rootstock, seedlings were grafted using the whip grafting method. In the third true leaf expansion stage, seedlings were foliar sprayed with Vc at concentrations of 50, 100, 150, and 200 mg L−1. Three days after initial spraying, seedlings were subjected to cold stress (8 °C) for 3 days, with continued spraying. After that, morphological and physiological parameters were assessed. Results showed that 150 mg L−1 Vc treatment was most impactive, significantly reducing the cold damage index while increasing the root-to-shoot ratio, root vitality, chlorophyll content, and activities of antioxidant enzymes (SOD, POD, CAT). Moreover, this treatment enhanced levels of soluble sugars, soluble proteins, and proline compared to control. However, 200 mg L−1 treatment elevated malondialdehyde (MDA) content, indicating potential oxidative stress. For transcriptomic analysis, leaves from the 150 mg L−1 Vc and CK treatments were sampled at 0, 1, 2, and 3 days of cold stress. Differential gene expression revealed that genes associated with photosynthesis (LHCA1), stress signal transduction (MYC2-1, MYC2-2, WRKY22, WRKY2), and antioxidant defense (SOD-1, SOD-2) were initially up-regulated and subsequently down-regulated, as validated by qRT-PCR. Overall, we found that the application of 150 mg L−1 Vc enhanced cold tolerance in grafted cucumber seedlings by modulating gene expression networks related to photosynthesis, stress response, and the antioxidant defense system. This study provides a way for developing Vc biostimulants to enhance cold tolerance in grafted cucumbers, improving sustainable cultivation in low-temperature regions. Full article
(This article belongs to the Section Plant Response to Abiotic Stress and Climate Change)
26 pages, 1260 KiB  
Article
Osteogenic Differentiation of Mesenchymal Stem Cells Induced by Geometric Mechanotransductive 3D-Printed Poly-(L)-Lactic Acid Matrices
by Harrison P. Ryan, Bruce K. Milthorpe and Jerran Santos
Int. J. Mol. Sci. 2025, 26(15), 7494; https://doi.org/10.3390/ijms26157494 (registering DOI) - 2 Aug 2025
Abstract
Bone-related defects present a key challenge in orthopaedics. The current gold standard, autografts, poses significant limitations, such as donor site morbidity, limited supply, and poor morphological adaptability. This study investigates the potential of scaffold geometry to induce osteogenic differentiation of human adipose-derived stem [...] Read more.
Bone-related defects present a key challenge in orthopaedics. The current gold standard, autografts, poses significant limitations, such as donor site morbidity, limited supply, and poor morphological adaptability. This study investigates the potential of scaffold geometry to induce osteogenic differentiation of human adipose-derived stem cells (hADSCs) through mechanotransduction, without the use of chemical inducers. Four distinct poly-(L)-lactic acid (PLA) scaffold architectures—Traditional Cross (Tc), Triangle (T), Diamond (D), and Gyroid (G)—were fabricated using fused filament fabrication (FFF) 3D printing. hADSCs were cultured on these scaffolds, and their response was evaluated utilising an alkaline phosphatase (ALP) assay, immunofluorescence, and extensive proteomic analyses. The results showed the D scaffold to have the highest ALP activity, followed by Tc. Proteomics results showed that more than 1200 proteins were identified in each scaffold with unique proteins expressed in each scaffold, respectively Tc—204, T—194, D—244, and G—216. Bioinformatics analysis revealed structures with complex curvature to have an increased expression of proteins involved in mid- to late-stage osteogenesis signalling and differentiation pathways, while the Tc scaffold induced an increased expression of signalling and differentiation pathways pertaining to angiogenesis and early osteogenesis. Full article
(This article belongs to the Special Issue Novel Approaches for Tissue Repair and Tissue Regeneration)
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