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16 pages, 13998 KB  
Article
Surfactin Inhibits Osteoclast Differentiation by Negatively Regulating the Elk1-AP-1-NFATc1 Axis
by Kazuki Maruyama, Ayaka Koga, Yuki Kodama, Ryota Yamasaki, Yoshie Nagai-Yoshioka, Jun J. Miyamoto, Kayoko Kuroishi, Kaori Gunjigake, Tatsuo Kawamoto and Wataru Ariyoshi
Biomedicines 2026, 14(1), 155; https://doi.org/10.3390/biomedicines14010155 - 11 Jan 2026
Viewed by 105
Abstract
Background/Objectives: Surfactin is a biosurfactant with various biological activities, including antibacterial and anti-inflammatory properties; however, its effects on bone metabolism remain poorly understood. This study aimed to investigate the effects of surfactin on osteoclast differentiation and elucidate its underlying molecular mechanisms. Methods [...] Read more.
Background/Objectives: Surfactin is a biosurfactant with various biological activities, including antibacterial and anti-inflammatory properties; however, its effects on bone metabolism remain poorly understood. This study aimed to investigate the effects of surfactin on osteoclast differentiation and elucidate its underlying molecular mechanisms. Methods: RAW264.7 cells were treated with receptor activator of nuclear factor-kappa B ligand (RANKL) and surfactin, and osteoclast differentiation and maturation were evaluated by tartrate-resistant acid phosphatase and F-actin staining, respectively. Gene expression of differentiation markers was assessed using real-time reverse transcription-quantitative polymerase chain reaction, while the kinetics of intracellular signaling molecules and transcription factors were analyzed using Western blot analysis. Results: Surfactin treatment significantly inhibited osteoclast differentiation and maturation, as well as the mRNA expression of Nfatc1, Acp5, and Cathepsin K. Although surfactin did not markedly affect RANKL-induced activation of the NF-κB or MAPK-mediated signaling, it significantly suppressed the expression of c-Fos at both the mRNA and protein levels. Furthermore, surfactin attenuated the phosphorylation of Elk1, a transcription factor involved in c-Fos induction. Conclusions: Surfactin inhibits RANKL-induced osteoclast differentiation by negatively regulating the Elk1-AP-1-NFATc1 axis. Surfactin may thus be a promising therapeutic candidate for the treatment of metabolic bone disorders and inflammatory bone destruction. Full article
(This article belongs to the Section Molecular and Translational Medicine)
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24 pages, 17450 KB  
Article
Integrated Single-Cell and Bulk Transcriptomics Unveils Immune Profiles in Chick Erythroid Cells upon Avian Pathogenic Escherichia coli Infection
by Fujuan Cai, Xianjue Wang, Chunzhi Wang, Yuzhen Wang and Wenguang Zhang
Animals 2026, 16(2), 179; https://doi.org/10.3390/ani16020179 - 7 Jan 2026
Viewed by 173
Abstract
Nucleated erythroid cells (NECs) have emerged as active participants in immune responses in addition to their canonical oxygen transport function. The subpopulations and immune heterogeneity of chick erythroid cells (ch-ECs) upon infection have not been fully characterized. Single-cell RNA sequencing (scRNA-seq) was used [...] Read more.
Nucleated erythroid cells (NECs) have emerged as active participants in immune responses in addition to their canonical oxygen transport function. The subpopulations and immune heterogeneity of chick erythroid cells (ch-ECs) upon infection have not been fully characterized. Single-cell RNA sequencing (scRNA-seq) was used to profile ch-ECs in chicks infected with avian pathogenic Escherichia coli (APEC). Unsupervised clustering uncovered ten distinct ch-EC subpopulations (C1–C10), with significant compositional shifts between infected and control groups. Pseudotime analysis revealed a developmental continuum: C1, C3, C5, and C9 as early progenitors; C2, C4, C6, C7, and C10 as mature erythroid cells; and C8 as a naive population. We revealed 62 immune-related genes, including protein kinases and heat shock proteins, and subpopulation-specific differentially expressed genes (DEGs) linked to immune functions. SCENIC analysis revealed Fos, Srf, and Stat3 as key transcription factors with elevated regulon activity and specificity following infection. Subpopulations C2, C4, C6, and C7, which exhibited marked abundance changes, were scrutinized for immune relevance through integrated multi-omics analysis. Immune-related genes including FOS, AKAP9, HS6ST1, GAB3, TFRC, HSPA8, HSP90AA1, and DNAJB6 were identified. Enrichment analysis indicated activation of the MHC class I antigen presentation pathway, while pathways such as Mitogen-Activated Protein Kinase (MAPK) signaling, NOD-like receptor (NLR) signaling, and the heat shock response were found to be suppressed. In conclusion, this study delineates the immune gene repertoire and signaling networks of ch-ECs during APEC infection, offering new perspectives on NEC immunoregulatory functions. Full article
(This article belongs to the Special Issue Bacterial Disease Research in Livestock and Poultry)
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25 pages, 1542 KB  
Article
Effect of a Supplement Containing Probiotics, Prebiotics, and Yeast Extract on Gut Inflammation, Microbiota, and Cytokines in Healthy Dogs
by Angela Zilinger, Mary K. Sramek, Tarun Chandra, Teresa Schmidt, Jessica Bagel, Andrew Stayduhar, James Fryer and Gregory D. Sunvold
Pets 2026, 3(1), 1; https://doi.org/10.3390/pets3010001 - 4 Jan 2026
Viewed by 775
Abstract
Probiotics, prebiotics, and postbiotics are of interest for their potential gastrointestinal and immunological benefits in pet health. This study aimed to assess whether a unique blend of Bacillus subtilis, Bacillus clausii, Bacillus coagulans (Weizmannia coagulans), FOS, GOS, and a [...] Read more.
Probiotics, prebiotics, and postbiotics are of interest for their potential gastrointestinal and immunological benefits in pet health. This study aimed to assess whether a unique blend of Bacillus subtilis, Bacillus clausii, Bacillus coagulans (Weizmannia coagulans), FOS, GOS, and a postbiotic yeast extract could provide beneficial gut and immunological effects when fed to healthy, adult dogs. Twenty-four healthy adult beagle dogs (mean age 5.17 yrs) were fed the probiotic, prebiotic, and yeast chew (PPYC) or control chew (CC) supplement for 31 days, accompanied by fecal and blood sampling. Following 31 days, PPYC fed dogs had decreased (p < 0.05) fecal calprotectin concentration, a biomarker indicative of reduced intestinal inflammation, compared with dogs receiving the CC. In the PPYC group, blood C-reactive protein levels, an indicator of tissue inflammation, tended (p = 0.11) to be reduced. In addition, dogs receiving the PPYC supplement showed an increase in the IL-17a cytokine (p < 0.05). Despite dogs being in a clinically healthy state, changes in some dysbiosis-related bacterial strains were observed. There was an increase (p < 0.05) in the % of total bacteria of Blautia in the PPYC group by the end of the study, as well as an increase in the percent change from Day 0 of C. hiranosis (p < 0.05). Increased alpha diversity, a measure related to the resilience to environmental change, was observed in the PPYC group (p < 0.05). These results suggest that after consuming a supplement containing probiotics, prebiotics and a postbiotic yeast extract, markers of gut and systemic health were improved in otherwise healthy dogs. Full article
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25 pages, 12788 KB  
Article
The Effect of Fructooligosaccharide and Inulin Addition on the Functional, Mechanical, and Structural Properties of Cooked Japonica Rice
by Bing Dai, Ruijun Chen, Shiyu Chang, Zheng Wei, Xiaohong Luo, Jiangzhang Wu and Xingjun Li
Gels 2026, 12(1), 48; https://doi.org/10.3390/gels12010048 - 1 Jan 2026
Viewed by 249
Abstract
To test whether fructooligosaccharide (FOS) and inulin (INU) molecules can improve the hardness of cooked rice through forming a hydrogel network, we added FOS or INU at 0%, 3%, 5%, 7%, and 10% concentrations to two cooking japonica rice and compared the cooking [...] Read more.
To test whether fructooligosaccharide (FOS) and inulin (INU) molecules can improve the hardness of cooked rice through forming a hydrogel network, we added FOS or INU at 0%, 3%, 5%, 7%, and 10% concentrations to two cooking japonica rice and compared the cooking and textural parameters, the pasting, thermal, and thermo-mechanical properties, and the microstructure of the cooked rice. General Linear Model Univariate (GLMU) analysis revealed that, compared with no oligofructose addition, both FOS and INU addition reduced the rice cooking time and increased the gruel solid loss. The addition of these dietary fibers (DFs) to cooking rice lowered the hardness, adhesiveness, springiness, gumminess, and chewiness of the rice, but maintained the cohesiveness and increased the resilience. Compared with no oligofructose addition, FOS and INU addition improved the smell, taste, and total sensory score of cooked rice. The addition of these DFs significantly decreased the trough, peak, final, breakdown, and setback viscosities, but increased the pasting temperature and peak time. Both FOS and INU addition decreased the enthalpy of gelatinization but increased the peak and conclusion temperature of gelatinization of rice flour paste. After the retrograded flour pastes were kept at 4 °C for 21 days, both FOS and INU significantly increased amylopectin aging compared with no oligofructose addition. The FOS-added and INU-added rice doughs had a higher dough development time and stability time, gelatinization peak torque, setback torque, and gelatinization speed, with a lower protein weakening degree, amylase activity, breakdown torque, heating speed, and enzymatic hydrolysis speed. Compared with no oligofructose addition, both FOS and INU addition reduced the amorphous region of starch and β-sheet percentage, but increased the percentages of random coils, α-helixes, and β-turns in cooked rice. Principal component analysis (PCA) further demonstrated that the gruel solid loss, cooked rice hardness, chewiness, gumminess, taste, and the peak, trough, breakdown, final, and setback viscosities were sensitive parameters for evaluating the effects of species and the amount of oligofructose addition on rice quality. The microstructure showed that FOS or INU addition induced thickening of the matrix walls and an increase in the pore size, forming a soft and evenly swollen structure. These results suggest that FOS or INU addition inhibits amylose recrystallization but maintains amylopectin recrystallization in cooked rice, with INU addition producing greater improvements in the texture and sensory scores of cooked rice compared withFOS addition. This study provides evidence of the advantages of adding DFs and probiotics such as INU and FOS to cooked rice. Full article
(This article belongs to the Special Issue Application of Composite Gels in Food Processing and Engineering)
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20 pages, 2158 KB  
Article
Dewatering of Sweet Whey Using Forward Osmosis on an Industrial Scale
by Bastian Greisner, Magnus Rienäcker, Frank Rögener and André Lerch
Purification 2026, 2(1), 1; https://doi.org/10.3390/purification2010001 - 26 Dec 2025
Viewed by 178
Abstract
Industrial whey dewatering via membrane processes remains challenging due to the rapid increase in viscosity, strong fouling tendencies from proteins and minerals, and the steep rise in osmotic pressure during concentration. These effects restrict operating windows and complicate energy-efficient process control. This study [...] Read more.
Industrial whey dewatering via membrane processes remains challenging due to the rapid increase in viscosity, strong fouling tendencies from proteins and minerals, and the steep rise in osmotic pressure during concentration. These effects restrict operating windows and complicate energy-efficient process control. This study addresses the application of forward osmosis (FO) technology for industrial-scale dewatering of sweet whey using an Aquaporin Inside® HFFO14 module. Various feed- and draw-side cross flow velocities (0.0397 to 0.0524 m s−1 and 0.0127 to 0.0190 m s−1, respectively) and draw solution (DS) osmotic pressures of 20 bar and 60 bar were investigated using a production-scale prototype plant. Sweet whey had an initial osmotic pressure of 7 bar and an electrical conductivity of 5.7 mS cm−1. DS pressures of 20 bar and 60 bar resulted in a total recovery of 50% and over 80%, respectively. Water flux rates initially ranged from 10.1 to 11.6 L m−2 h−1 (LMH) and ceased at 3.3 LMH. Specific energy demand ranged from 0.15 to 1.1 kWh m−3. These findings support the feasibility of industrial-scale FO technology and underscore the potential of FO as an energy-efficient, sustainable solution for the dairy industry. However, frequent rinsing and cleaning routines are crucial to maintain membrane performance. Full article
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19 pages, 1968 KB  
Article
Proteomic Analysis of Streptococcus suis During Exposure to Intracellular Condition of Human Macrophage U937 Cells
by Peerarin Prangsuwan, Orathai Yinsai, Sittiruk Roytrakul and Kwanjit Duangsonk
Int. J. Mol. Sci. 2026, 27(1), 128; https://doi.org/10.3390/ijms27010128 - 22 Dec 2025
Viewed by 343
Abstract
Streptococcus suis is an important zoonotic pathogen responsible for severe infections in pigs and humans. Its capacity to survive within phagocytic cells is considered a key virulence mechanism that contributes to dissemination and persistence in host tissues. This study employed comparative proteomic profiling [...] Read more.
Streptococcus suis is an important zoonotic pathogen responsible for severe infections in pigs and humans. Its capacity to survive within phagocytic cells is considered a key virulence mechanism that contributes to dissemination and persistence in host tissues. This study employed comparative proteomic profiling to investigate intracellular adaptation of S. suis serotypes 2 (SS2) and 14 (SS14) during infection of human U937 macrophages. Five isolates originating from humans and pigs were analyzed using gel electrophoresis with liquid chromatography–tandem mass spectrometry (GeLC–MS/MS), revealing 118 differentially expressed proteins grouped into 11 functional categories. Translation-related proteins represented the largest group (48%), including upregulated ribosomal subunits (30S: S2, S5, S7, S8, S12, S15; 50S: L1, L5, L18, L22, L24, L33, L35) and translation factors such as GidA/TrmFO and RimP. Enrichment of carbohydrate metabolism and DNA replication proteins, including phosphoenolpyruvate carboxylase (PEP), UDP-N-acetylglucosamine pyrophosphorylase (GlmU), and ATP-dependent DNA helicase RuvB, indicated metabolic reprogramming and stress adaptation under intracellular conditions. Stress-response proteins such as molecular chaperone DnaK were also induced, supporting their multifunctional, “moonlighting” roles in virulence and host interaction. Comparative analysis showed that SS2 expressed a broader range of adaptive proteins than SS14, consistent with its higher virulence potential. These findings reveal conserved intracellular responses centered on translation, energy metabolism, and stress tolerance, which enable S. suis to survive within human macrophages. Integration of these intracellular proteomic signatures with previous exoproteomic, peptidomic, and network-based studies highlights translational and metabolic proteins—particularly DnaK, enolase, elongation factor EF-Tu, and GlmU—as multifunctional candidates linking survival and immunogenicity. This work establishes a comparative proteomic foundation for understanding S. suis intracellular adaptation and highlights potential targets for future vaccine or therapeutic development against this zoonotic pathogen. Full article
(This article belongs to the Section Molecular Informatics)
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21 pages, 10395 KB  
Article
Aqueous Extract of Fructus Choerospondiatis Peel Suppresses Vascular Inflammation and Alleviates Atherosclerosis via AKT/c-FOS/IL-6 Axis
by Andong Wu, Jiayi Dong, Jiankun Liu, Xueting Gong, Xueer Li, Bingbing Zhou, Ming Wan, Weixin Lv, Jiayu Qiu, Ya Zhao, Yu Fang, Jie Huang and Xiao-Li Tian
Nutrients 2026, 18(1), 21; https://doi.org/10.3390/nu18010021 - 19 Dec 2025
Viewed by 345
Abstract
Background: Atherosclerosis is the pathological basis for lethal cardio-cerebral vascular diseases, such as coronary artery disease and stroke. Fructus Choerospondiatis (FC) has demonstrated cardiac protective effects in multiple ethnomedicine. Whether these protective effects are attributed to the prevention of vascular atherosclerosis, however, [...] Read more.
Background: Atherosclerosis is the pathological basis for lethal cardio-cerebral vascular diseases, such as coronary artery disease and stroke. Fructus Choerospondiatis (FC) has demonstrated cardiac protective effects in multiple ethnomedicine. Whether these protective effects are attributed to the prevention of vascular atherosclerosis, however, remains unknown. We aim to examine the anti-atherosclerotic effect of FC aqueous extract and elucidate the underlying mechanism. Methods: FC was separated into peel and pulp, and the aqueous extract was obtained separately by boiling in water to mimic decocting. Atherosclerosis model was established in ApoE−/− mice fed with a high-fat diet, and histological analysis were utilized to evaluate the development of atherosclerosis. Various inflammatory models were constructed in mice to evaluate the anti-inflammatory effect of FC extract systemically, including acute local inflammation induced by traumatic injury (ear/foot swelling), acute systemic inflammation triggered by pathogenic infection (LPS- and POLY (I:C)-induced), as well as chronic inflammatory conditions associated with oxidative stress (D-galactose-induced), metabolic disorder (db/db mice), and aging. LC-MS and network pharmacology identified bioactive components and targets. Western blotting, ELISA, qPCR, and immunofluorescence were utilized to analyze the key genes involved in the mechanisms. Results: FC peel extract reduced serum IL-6 level, atherosclerotic plaque area, and macrophage content in the plaque, while pulp extract showed no protective effects. Peel extract exhibits anti-inflammatory effects in all models. The integrative application of LC-MS and network pharmacology identified ellagic acid as the major bioactive component and AKT as its target protein. Mechanistically, FC peel extract inhibits AKT phosphorylation, suppresses c-FOS expression and nuclear translocation, reduces IL-6 transcription and inflammation, and thus alleviates atherosclerosis. Conclusions: FC peel aqueous extract exerts anti-atherosclerotic effect by inhibiting inflammation through AKT/c-FOS/IL-6 axis. This study provides novel insights into the protective effects against atherosclerosis of FC peel and highlights its potential application in the prevention and treatment of coronary artery diseases. Full article
(This article belongs to the Special Issue Preventive and Therapeutic Nutraceuticals)
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19 pages, 3961 KB  
Article
Retinal Degeneration in Alzheimer’s Disease 5xFAD Mice Fed DHA-Enriched Diets
by Mário S. Pinho, Husaifa Ahfaz, Sandra Carvalho, Jorge Correia, Maria Spínola, José M. Pestana, Narcisa M. Bandarra and Paula A. Lopes
Cells 2026, 15(1), 8; https://doi.org/10.3390/cells15010008 - 19 Dec 2025
Viewed by 558
Abstract
Alzheimer’s disease (AD) is marked by cognitive decline, and also by retinal degeneration. Having in mind that docosahexaenoic acid (DHA, 22:6n − 3) is a safe, low-cost, and pivotal fatty acid for brain health and sustained cognitive function, this study exploits environmentally friendly [...] Read more.
Alzheimer’s disease (AD) is marked by cognitive decline, and also by retinal degeneration. Having in mind that docosahexaenoic acid (DHA, 22:6n − 3) is a safe, low-cost, and pivotal fatty acid for brain health and sustained cognitive function, this study exploits environmentally friendly non-fish sources as potential dietary supplements enriched with DHA to prevent or reverse AD. Forty 5xFAD transgenic male mice, aged five weeks old, were randomly distributed by five body weight-matched dietary groups (with eight animals each) and fed isocaloric diets based on the AIN-93M standard formulation for rodents for 6 months. Except for the control feed (without supplementation), each diet contained a modified lipidic fraction supplemented with 2% of the following: (1) linseed oil (LSO, rich in alpha-linolenic acid (ALA, 18:3n − 3)); (2) cod liver oil (fish oil, FO, rich in both DHA and eicosapentaenoic acid (EPA, 20:5n − 3)); (3) Schizochytrium sp. microalga oil (Schizo, with 40% of DHA); and (4) commercial DHASCO (DHASCO, with 70% of DHA). The aim of this study was to measure retinal neural layer thickness, calculate ganglion cell layer (GCL) density, and assess retinal injury by means of immunohistochemical staining for β-amyloid plaques deposition, TAU protein levels, and IBA1, as hallmark features of AD progression, in order to elucidate the effects of different dietary DHA treatments in Alzheimer’s retinas. Although no statistical differences were observed across retinal layer thicknesses depending on the diet (p > 0.05), there was a consistent pattern for slightly increased retinal thickness in 5xFAD mice fed fish oil relative to the others for the measurement of total layers, in general and for the inner segment/outer segment layer, the outer nuclear layer, the outer plexiform layer, the inner nuclear layer, and the inner plexiform layer, in particular. The ganglion cell layer (GCL) density was increased in 5xFAD mice fed the DHASCO oil diet relative to the control (p < 0.05), suggesting a benefit of DHA supplementation on the number of viable ganglion cells. No positive staining was observed for β-amyloid plaques deposition or the neuroinflammatory marker, IBA1, corroborating previous findings in human AD retinas. Conversely, the internal retinal layers showed intense TAU immunostaining. Immnunostained TAU area was significantly reduced in 5xFAD mice fed a fish oil diet compared to control (p < 0.05), although the number of TAU-positive cells did not differ across diets (p > 0.05). The retinal protected integrity derived from the benefits of DHA supplementation found, either from fish oil or DHASCO oil, underscores the potential of retinal biomarkers as non-invasive indicators of cognitive decline and overall brain health, opening new avenues for investigating AD pathophysiology in the retina. Full article
(This article belongs to the Special Issue Advances in the Discovery of Retinal Degeneration)
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17 pages, 2646 KB  
Article
Establishment of an Isolation System for Extracellular Vesicles of Fusarium oxysporum and Its Proteomic Analysis
by Jiayi Lou, Guangjin Hu, Xuan Wang, Qiang Liu, Yuwei Chen and Weichun Zhao
J. Fungi 2025, 11(12), 884; https://doi.org/10.3390/jof11120884 - 15 Dec 2025
Viewed by 534
Abstract
Extracellular vesicles (EVs) secreted by Fusarium oxysporum play an important role in the process of its infestation of the host, but the in vitro research system for EVs of F. oxysporum (Fo-EVs) has not yet been improved, and the mechanism of [...] Read more.
Extracellular vesicles (EVs) secreted by Fusarium oxysporum play an important role in the process of its infestation of the host, but the in vitro research system for EVs of F. oxysporum (Fo-EVs) has not yet been improved, and the mechanism of its action remains unclear. In this study, particle size distribution, particle concentration, number of particles per unit of protein, number of particles per unit of mycelial biomass, and concentration of contaminated proteins were used as indicators to evaluate the yield and purity of Fo-EVs. The optimal method for Fo-EV preparation and extraction was screened by comparing liquid culture, solid culture, and solid culture with enzymatic cell wall hydrolysis. The optimal system for Fo-EVs separation and purification was screened by a pairwise combination of three primary methods (Ultracentrifugation (UC), Ultrafiltration (UF), and Polyethylene glycol precipitation method (PEG)) and two secondary methods (Size-exclusion chromatography (SEC) and Aqueous two-phase system (ATPS)), respectively. The protein composition was identified via mass spectrometry technology, followed by GO annotation and GO enrichment analysis using whole-genome proteins as the background. Based on these steps, a Fo-EV protein library was constructed to reveal Fo-EV’s most active biological functions. The results showed that solid culture combined with the UC-SEC method could effectively enrich Fo-EVs with a typical cup-shaped membrane structure. The obtained Fo-EVs had an average particle size of 253.50 nm, a main peak value of 200.60 nm, a particle concentration of 2.04 × 1010 particles/mL, and a particle number per unit protein of 1.09 × 108 particles/μg, which were significantly superior to those of other combined methods. Through proteomic analysis, 1931 proteins enriched in Fo-EVs were identified, among which 350 contained signal peptides and 375 had transmembrane domains. GO enrichment analysis revealed that these proteins were mainly involved in cell wall synthesis, vesicle transport, and pathogenicity-related metabolic pathways. Additionally, 9 potential fungal EV markers, including Hsp70, Rho GTPase family, and SNARE proteins, were screened. This study constructed an isolation system and a marker database for Fo-EVs, providing a methodological and theoretical basis for in-depth analysis of the biological functions of Fo-EVs. Full article
(This article belongs to the Special Issue Fungal-Related Proteomics in Biotechnology and Health)
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18 pages, 3432 KB  
Article
Kölliker–Fuse/Parabrachial Complex PACAP—Glutamate Pathway to the Extended Amygdala Couples Rapid Autonomic and Delayed Endocrine Responses to Acute Hypotension
by Vito S. Hernández, Pedro Segura-Chama and Limei Zhang
Int. J. Mol. Sci. 2025, 26(23), 11405; https://doi.org/10.3390/ijms262311405 - 25 Nov 2025
Viewed by 421
Abstract
The calyx of Held is a giant axo-somatic synapse classically confined to the auditory brainstem. We recently identified morphologically similar calyx-like terminals in the extended amygdala (EA) that arise from the ventrolateral parabrachial complex and co-express PACAP, CGRP, VAChT, VGluT1, and VGluT2, targeting [...] Read more.
The calyx of Held is a giant axo-somatic synapse classically confined to the auditory brainstem. We recently identified morphologically similar calyx-like terminals in the extended amygdala (EA) that arise from the ventrolateral parabrachial complex and co-express PACAP, CGRP, VAChT, VGluT1, and VGluT2, targeting PKCδ+/GluD1+ EA neurons. Here, we asked whether this parabrachial–EA pathway participates in compensation during acute hypotension. In rats given hydralazine (10 mg/kg, i.p.), we quantified Fos protein during an early phase (60 min) and a late phase (120 min). Early after hypotension, Fos surged in a discrete subpopulation of the parabrachial Kölliker–Fuse (KF) region and in the EA, whereas magnocellular neurons of the supraoptic and paraventricular nuclei (SON/PVN) remained largely silent. By 120 min, magnocellular SON/PVN neurons were robustly Fos-positive. Confocal immunohistochemistry showed that most Fos+ PKCδ+/GluD1+ EA neurons were encircled by PACAP+ perisomatic terminals (80.8%), of which the majority co-expressed VGluT1 (88.1%). RNAscope in situ hybridization further identified a selective KF population co-expressing Adcyap1 (PACAP) and Slc17a7 (VGluT1) that became fos-positive during the early phase. Together, these data suggest that a KF PACAP+/VGluT1+ projection forms calyceal terminals around PKCδ+/GluD1+ EA neurons, providing a high-fidelity route for rapid autonomic rebound to falling blood pressure, while slower endocrine support is subsequently recruited via neurohormone-magnocellular activation. This work links multimodal parabrachial output to temporally layered autonomic–neuroendocrine control. Full article
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6 pages, 443 KB  
Proceeding Paper
Preliminary Studies on the Biosynthesis of Microbial Inulinase by Aspergillus niger ICCF 92
by Mariana Gratiela Vladu, Mihaela Carmen Eremia, Dana Maria Miu, Gabriela Valeria Savoiu and Maria Monica Petrescu
Chem. Proc. 2025, 18(1), 48; https://doi.org/10.3390/ecsoc-29-26690 - 11 Nov 2025
Viewed by 116
Abstract
Modern diets are high in fructans, which may lead to abdominal discomfort, particularly in sensitive individuals. Microbial inulinase, an enzyme that hydrolyzes inulin into fructose and fructo-oligosaccharides (FOS), has significant prebiotic potential and may contribute to the prevention of metabolic disorders by enhancing [...] Read more.
Modern diets are high in fructans, which may lead to abdominal discomfort, particularly in sensitive individuals. Microbial inulinase, an enzyme that hydrolyzes inulin into fructose and fructo-oligosaccharides (FOS), has significant prebiotic potential and may contribute to the prevention of metabolic disorders by enhancing fructan digestion. This study investigates inulinase production by the Aspergillus niger ICCF 92 strain under various growth conditions. Three carbon sources (inulin, molasses, and carob pod decoction), the time required for biosynthesis processes, and stirring speed were evaluated for their influence on inulinase activity. Nitrogen sources included yeast extract, ammonium nitrate, and ammonium phosphate. Process monitoring included pH measurement, protein quantification via the Bradford assay, and inulinase activity assessment using the 3,5-dinitrosalicylic acid method. The highest inulinase production (38.29 U/mL) and protein concentration (0.7548 mg/mL) were achieved after 14 days of static fermentation with carob pod decoction as the carbon source. Full article
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15 pages, 866 KB  
Article
Investigation of Listeria monocytogenes in Food in Northwestern Italy (2020–2024)
by Monica Pitti, Matteo Tavecchia, Angelo Romano, Simona Carrella, Giovanna Previto and Daniela Manila Bianchi
Foods 2025, 14(21), 3788; https://doi.org/10.3390/foods14213788 - 5 Nov 2025
Viewed by 1109
Abstract
Listeria monocytogenes is a foodborne pathogen of significant public health concern due to its high environmental resilience and ability to cause severe infections in vulnerable populations. The objective of the present study is to characterize foodborne strains of Listeria monocytogenes isolated between 2020 [...] Read more.
Listeria monocytogenes is a foodborne pathogen of significant public health concern due to its high environmental resilience and ability to cause severe infections in vulnerable populations. The objective of the present study is to characterize foodborne strains of Listeria monocytogenes isolated between 2020 and 2024 in northwestern Italy. Lm was detected through isolation, biochemical confirmation and molecular serogrouping. Next generation sequencing (NGS) analysis was used to characterize the strains in terms of virulence and antibiotic resistance. A total of 39 positive samples were identified from various food matrices, including meat products, fish, cheeses and ready-to-eat foods. The most frequently detected serogroups were IIc and IIa, with a notable presence of the highly virulent IVb group. Next-generation sequencing (NGS) was applied to all isolates, revealing the presence of virulence genes associated with the LIPI-1 island and internalins. In addition to pathogenicity islands, genes related to stress resistance (clpCEP, Gad A, GadB, GadC), biofilm production (agrA, flaA, degU, hfq) and sortase-mediated anchoring of surface protein (strA, strB) have been identified. The presence of antibiotic resistance genes was confirmed, with all isolates harboring the fosX gene. Moreover, four isolates exhibited resistance determinants against antibiotics belonging to two different classes: tetracyclines (tetM) and lincosamides (lsa(A)). Multilocus sequence typing (MLST) showed that clonal complex CC9 was the most prevalent among the isolates. Further, cgMLST and SNP analyses identified a principal cluster of closely related strains, which were isolated from meat products. These findings highlight the need for continuous surveillance of L. monocytogenes. Full article
(This article belongs to the Special Issue Detection and Control of Foodborne Pathogens in Food Supply Chain)
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18 pages, 1749 KB  
Article
Effect of Nitric Oxide on Adventitious Root Development from Cuttings of Sweetpotato and Associated Biochemical Changes
by Meng Wang, Jianghui Li, Yuhao Wu, Hongxing Zhang, Hui Wang and Lingyun Wang
Plants 2025, 14(20), 3183; https://doi.org/10.3390/plants14203183 - 16 Oct 2025
Viewed by 707
Abstract
Adventitious rooting is a key step for the clonal propagation of many economically important horticultural and woody species. Accumulating evidence suggests that nitric oxide (NO) serves as a key signaling molecule with key roles in root organogenesis. However, the role of NO in [...] Read more.
Adventitious rooting is a key step for the clonal propagation of many economically important horticultural and woody species. Accumulating evidence suggests that nitric oxide (NO) serves as a key signaling molecule with key roles in root organogenesis. However, the role of NO in adventitious root development and its underlying mechanism in sweetpotato cuttings remain to be clarified. In this study, a pot experiment was conducted using hydroponically cultured sweetpotato cuttings (Ipomoea batatas cv. ‘Jin Ganshu No. 9’) treated with different concentrations of sodium nitroprusside (SNP, an NO donor) solution (0, 10, 50, 100, 200, and 500 μmol·L−1). Three treatments were established: Control, SNP (the optimal concentration of SNP), and SNP + 2-(4-carboxyphenyl)-4,4,5,5-tetramethylimidazoline-1-oxyl-3-oxide (cPTIO, an NO scavenger). The results showed that NO promoted adventitious rooting in a dose-dependent manner, with the maximal biological response observed at 100 μM SNP. At this concentration, the root number and length of adventitious roots increased by 1.22 and 2.36 times, respectively, compared to the control. SNP treatment increased fresh root weight, dry root weight, the content of soluble sugar, soluble protein, chlorophyll a (Chl a), chlorophyll b (Chl b), and total chlorophyll (a + b) [Chl(a + b)], as well as the activities of peroxidase (POD), polyphenol oxidase (PPO), and indole acetic acid oxidase (IAAO). It also enhanced the levels of maximum fluorescence (Fm), maximum photochemical efficiency of photosystem II (Fv/Fm), absorbed light energy (ABS/RC), trapped energy flux (TRo/RC), and electron transport flux (ETo/RC), while decreasing starch content and initial fluorescence (Fo). On the 7th day, the SNP treatment significantly enhanced several biochemical parameters compared to the control. We observed an increase in many of the parameters: POD activity by 1.35 times, PPO activity by 0.55 times, chlorophyll content (Chl a by 0.66 times, Chl b by 0.22 times, and Chl a + b by 0.57 times), and photosynthesis parameters by 28–98%. Meanwhile, starch content and Fo in the SNP treatment decreased by 10.77% and 23.86%, respectively, compared to the control. Furthermore, the positive effects of NO on adventitious root development and associated biochemical parameters were reversed by the NO scavenger cPTIO. Additionally, significant and positive correlations were observed between morphological characteristics and most physiological indicators. Collectively, these results demonstrate that NO promotes adventitious root formation, which may be by enhancing rooting-related enzyme activities, improving photosynthetic performance in leaves, and accelerating the metabolism of soluble sugar, soluble protein, and starch. Full article
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36 pages, 7997 KB  
Article
The Cannabinoid CB1 Receptor Inverse Agonist/Antagonist SR141716A Activates the Adenylate Cyclase/PKA Signaling Pathway Among Other Intracellular Emetic Signals to Evoke Vomiting in Least Shrews (Cryptotis parva)
by Yina Sun, Louiza Belkacemi, Weixia Zhong, Zollie Daily and Nissar A. Darmani
Int. J. Mol. Sci. 2025, 26(20), 9884; https://doi.org/10.3390/ijms26209884 - 11 Oct 2025
Viewed by 794
Abstract
Intracellular emetic signals involved in the cannabinoid CB1 receptor inverse agonist/antagonist SR141716A were investigated. SR141716A (20 mg/kg, i.p.)-evoked vomiting occurred via both the central and peripheral mechanisms. This was accompanied by robust emesis-associated increases in the following: (i) c-fos- and [...] Read more.
Intracellular emetic signals involved in the cannabinoid CB1 receptor inverse agonist/antagonist SR141716A were investigated. SR141716A (20 mg/kg, i.p.)-evoked vomiting occurred via both the central and peripheral mechanisms. This was accompanied by robust emesis-associated increases in the following: (i) c-fos- and phospho-glycogen synthase kinase-3α/β (p-GSK-3αβ)-expression in the shrew’s dorsal vagal complex (DVC), (ii) phospho-extracellular signal-regulated kinase1/2 (p-ERK1/2) expression in both the DVC and jejunal enteric nervous system, and (iii) time-dependent upregulation of cAMP levels and phosphorylation of protein kinase A (PKA), protein kinase B (Akt), GSK-3α/β, ERK1/2, and protein kinase C αβII (PKCαβII) in the brainstem. SR141716A-evoked emetic parameters were attenuated by diverse inhibitors of the following: PKA, ERK1/2, GSK-3, phosphatidylinositol 3-kinase (PI3K)-Akt pathway, phospholipase C (PLC), PKC, Ca2+/calmodulin-dependent protein kinase II (CaMKII), L-type Ca2+ channel (LTCC), store-operated Ca2+ entry (SOCE), inositol trisphosphate receptor (IP3R), ryanodine receptor (RyRs), both 5-HT3-, and D2/3-receptor antagonists, and the transient receptor potential vanilloid 1 receptor (TRPV1R) agonist. SR141716A appears to evoke vomiting via inverse agonist activity involving emesis-associated kinases, including cAMP/PKA, ERK1/2, PI3K/Akt/GSK-3, PLC/PKCαβII, and CaMKII, which depend upon Ca2+ mobilization linking extracellular Ca2+ entry via plasma membrane Ca2+ channels (LTCC, SOCE, TRIPV1R) and intracellular Ca2+ release via IP3Rs and RyRs. The 5-HT3, NK1, and D2/3 receptors also contribute to SR141716A-mediated vomiting. Full article
(This article belongs to the Special Issue G Protein-Coupled Receptors)
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13 pages, 3209 KB  
Article
Fabrication and Measurement of Fiber Optic Sensor Based on Localized Surface Plasmon Resonance for Interleukin-8 Detection Using Micropillar and Gold Nanoparticle Composite
by Min-Jun Kim, Jong-Hyun Bang, Hyeong-Min Kim, Jae-Hyoung Park and Seung-Ki Lee
Appl. Sci. 2025, 15(20), 10894; https://doi.org/10.3390/app152010894 - 10 Oct 2025
Viewed by 1060
Abstract
This study reports the development of a fiber-optic localized surface plasmon resonance (FO-LSPR) sensor incorporating a three-dimensional micropillar array functionalized with gold nanoparticles. The micropillar structures were fabricated on the fiber facet using a single-mask imprint lithography process, followed by nanoparticle immobilization to [...] Read more.
This study reports the development of a fiber-optic localized surface plasmon resonance (FO-LSPR) sensor incorporating a three-dimensional micropillar array functionalized with gold nanoparticles. The micropillar structures were fabricated on the fiber facet using a single-mask imprint lithography process, followed by nanoparticle immobilization to create a composite plasmonic surface. Compared with flat polymer-coated fibers, the micropillar array markedly increased the effective sensing surface and enhanced light trapping by providing anti-reflective conditions at the interface. Consequently, the sensor demonstrated superior performance in refractive index sensing, yielding a sensitivity of 4.54 with an R2 of 0.984, in contrast to 3.13 and 0.979 obtained for the flat counterpart. To validate its biosensing applicability, Interleukin-8 (IL-8), a cancer-associated cytokine, was selected as a model analyte. Direct immunoassays revealed quantitative detection across a broad dynamic range (0.1–1000 pg/mL) with a limit of detection of 0.013 pg/mL, while specificity was confirmed against non-target proteins. The proposed FO-LSPR platform thus offers a cost-effective and reproducible route to overcome the surface-area limitations of conventional designs, providing enhanced sensitivity and stability. These results highlight the potential of the micropillar-based FO-LSPR sensor for practical deployment in point-of-care diagnostics and real-time biomolecular monitoring. Full article
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