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23 pages, 7458 KB  
Article
High-Glucose-Induced Metabolic and Epithelial Stress in Grass Carp Intestinal Epithelial Cells Associated with Methylation-Related Transcriptional Responses
by Linjie Qian, Wenqiang Jiang, Yan Lin, Siyue Lu, Xianping Ge and Linghong Miao
Int. J. Mol. Sci. 2026, 27(13), 5732; https://doi.org/10.3390/ijms27135732 - 25 Jun 2026
Viewed by 317
Abstract
High-glucose exposure impairs intestinal metabolic homeostasis and barrier integrity in fish, but the transcriptional responses associated with high-glucose adaptation in fish intestinal epithelial cells remain incompletely understood. This study investigated whether exogenous 5-methylcytosine (5MC) alleviates high-glucose-induced metabolic and epithelial stress in grass carp [...] Read more.
High-glucose exposure impairs intestinal metabolic homeostasis and barrier integrity in fish, but the transcriptional responses associated with high-glucose adaptation in fish intestinal epithelial cells remain incompletely understood. This study investigated whether exogenous 5-methylcytosine (5MC) alleviates high-glucose-induced metabolic and epithelial stress in grass carp (Ctenopharyngodon Idella) intestinal epithelial cells and whether these responses are associated with changes in DNA methyltransferase 3 beta (dnmt3b) expression and Caudal type homeobox 1b (cdx1b)/Sodium-glucose cotransporter 1 (sglt1)-related transcriptional responses. As exploratory in silico information, molecular docking predicted candidate complex conformations of DNMT3B with CDX1B and SGLT1, with binding energies of −37.2 and −25.9 kcal/mol, respectively. Functionally, dnmt3b knockdown significantly reduced dnmt3b, Interleukin 6 (il6), and Nuclear factor kappa B (nfκb) expression, while increasing cdx1b, sglt1, Solute carrier family 2 member 3a (slc2a3a), 6-Phosphofructo-2-kinase/fructose-2,6-bisphosphatase 4a (pfkfb4a), and Amine oxidase copper containing 1 (aoc1) expression (p < 0.05). CDX2/CDX1B-like immunoreactive protein and SGLT1 protein levels were also increased after dnmt3b knockdown (p < 0.05). Under high-glucose stress, exogenous 5MC exerted concentration-dependent effects. Specifically, 6 mM 5MC significantly reduced residual extracellular glucose, lactate dehydrogenase and diamine oxidase activities, and malondialdehyde content, while increasing glutathione content, cell viability, and cell migration (p < 0.05). These effects remained detectable after replacement with high-glucose medium for an additional 12 h. By contrast, 24 mM 5MC markedly increased lactate dehydrogenase activity and reduced cell viability, suggesting potential cytotoxicity (p < 0.05). S-adenosylmethionine (SAM) levels were significantly lower in the NC and 6 mM groups than in the HG, 12 mM, and 24 mM groups, suggesting changes in SAM-related one-carbon metabolic status rather than direct evidence of altered DNA methylation (p < 0.05). Exogenous 5MC, particularly at 6 mM, alleviated high-glucose-induced metabolic and epithelial stress in grass carp intestinal epithelial cells. These effects were accompanied by changes in several glucose metabolism- and inflammation-related genes. However, the cellular uptake, metabolic fate, DNA incorporation, methylation consequences, and causal roles of these gene-expression changes remain to be further verified. Full article
(This article belongs to the Special Issue The Latest Molecular Insights into Animal Nutrition)
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18 pages, 6886 KB  
Article
Spiny Mice Show a Profibrotic Epicardial Mesothelial Response to Hypoxic Injury Comparable to C57BL/6 Mice
by Konstantin Dergilev, Aleria Dolgodvorova, Zoya Tsokolaeva, Irina Iarushkina, Irina Beloglazova, Yulia Goltseva and Yelena Parfyonova
Biomolecules 2026, 16(5), 717; https://doi.org/10.3390/biom16050717 - 13 May 2026
Viewed by 612
Abstract
Epicardial mesothelium plays a pivotal role in postinfarction cardiac repair by generating fibroblasts, producing extracellular matrix, and releasing paracrine mechanisms. However, interspecies differences have not been sufficiently studied, particularly in in vivo models of scar-free healing such as the African spiny mouse ( [...] Read more.
Epicardial mesothelium plays a pivotal role in postinfarction cardiac repair by generating fibroblasts, producing extracellular matrix, and releasing paracrine mechanisms. However, interspecies differences have not been sufficiently studied, particularly in in vivo models of scar-free healing such as the African spiny mouse (Acomys cahirinus). This study aimed to compare the profibrotic response of epicardial mesothelial cells (MCs) from Acomys and C57BL/6 mice to hypoxic stress, a key factor in postinfarction recovery. We isolated epicardial MCs from the African spiny mouse (Acomys cahirinus), a species with documented cardiac regenerative capabilities, and from C57BL/6 laboratory mice. Using a CoCl2-induced hypoxia model in vitro, we assessed cell viability, morphological changes, and expression of epithelial and fibroblast markers. In vivo, following experimental myocardial infarction (MI), we evaluated tissue hypoxia (pimonidazole adducts), epicardial activation (layer thickness, Wt1+ and TBX18+ progenitor cells), and collagen accumulation. The study was conducted using real-time PCR, Western blotting, immunohistochemical analysis and microscopic examination. In vitro, MCs from both species exhibited an epithelial-like phenotype under normoxic conditions, expressing E-cadherin and cytokeratin 18. Hypoxia (200 µM CoCl2) induced a comparable response in both Acomys and C57BL/6 cells, characterized by a shift to a spindle-shaped, fibroblast-like morphology, decreased E-cadherin expression, and increased pro-collagen 1 and α-SMA expression. Following MI, both species exhibited similarly extensive hypoxic areas affecting the epicardial zone. Epicardial activation dynamics were comparable: from day 3 post-MI, epicardial thickness increased significantly, and Wt1+ and TBX18+ progenitor cells accumulated, peaking during the first week. Collagen accumulation in the epicardial region was similar between species, although the number of Wt1+ cells was higher in C57BL/6 on day 7. Despite the well-known superior regenerative capacity of spiny mice, epicardial MCs from Acomys and C57BL/6 demonstrated similar signs of profibrotic responses to hypoxic stimulation both in vitro and following MI. These findings suggest that species-specific regenerative outcomes may not be attributable to differential acute epicardial sensitivity to hypoxia, but rather to downstream mechanisms or additional factors influencing the cardiac repair process. This study provides the first characterization of Acomys epicardial MCs and establishes a foundation for further investigation of evolutionarily conserved and species-specific mechanisms of cardiac regeneration. Full article
(This article belongs to the Special Issue New Insights into Mesothelial Cells)
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27 pages, 3102 KB  
Article
Proton Irradiation Induces Differential Cellular Responses and Proteomic Signatures in Chondrosarcoma and Chondrocytes
by Mihaela Tudor, Roxana Cristina Popescu, Benoît Bernay, Mihaela Temelie, Liviu Craciun, Tiberiu Relu Esanu, François Chevalier and Diana Iulia Savu
Curr. Issues Mol. Biol. 2026, 48(5), 450; https://doi.org/10.3390/cimb48050450 - 25 Apr 2026
Viewed by 488
Abstract
Chondrosarcoma (CHS), the second most common primary malignant cartilage tumor, is largely resistant to conventional therapies, making surgical resection the standard treatment. Proton therapy offers a physical advantage through the Bragg peak, enabling targeted irradiation while sparing surrounding tissues. However, differential biological responses [...] Read more.
Chondrosarcoma (CHS), the second most common primary malignant cartilage tumor, is largely resistant to conventional therapies, making surgical resection the standard treatment. Proton therapy offers a physical advantage through the Bragg peak, enabling targeted irradiation while sparing surrounding tissues. However, differential biological responses between malignant and normal cartilage cells remain poorly understood. In this study, CHS SW1353 cells and normal chondrocytes (MC615) were exposed to proton irradiation. Biological responses were assessed via clonogenic survival, cell viability, apoptosis (caspase 3/7), micronucleus formation, cell cycle profiling, and oxidative stress markers. Proteomic changes were analyzed using mass spectrometry and bioinformatics. CHS cells exhibited higher radioresistance (D10 = 6.45 Gy) than normal chondrocytes (D10 = 5.08 Gy), oxidative stress adaptation, G1 arrest and proteomic plasticity, whereas normal chondrocytes displayed increased oxidative stress, extracellular matrix fragility and impaired integrin signaling. Notably, the tumor-specific increased levels of Tyrosine-protein kinase Fyn and Yes1-associated transcriptional regulator (YAP1) signaling suggest molecular drivers of radioresistance. Overall, proton irradiation elicits distinct biological and proteomic responses in malignant versus normal cartilage cells. These findings highlight potential radiosensitization targets, including Fyn/Src and YAP1/Hippo pathways, while underscoring the need to optimize proton therapy to enhance tumor control while minimizing damage to healthy cartilage. Full article
(This article belongs to the Special Issue Radiation-Induced Cellular and Molecular Responses)
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21 pages, 4584 KB  
Article
Interaction Between Telocytes and Mast Cells in Genetically Determined Non-Obstructive Azoospermia with AZFc Deletion: An Ultrastructural Study
by Irina Chekmareva, Andrey Kostin, Nina Kulchenko, Grigory Demyashkin, Oksana Paklina, Alexander Alekhnovich, Artem Volodkin, Atim Emaimo John, Ilya Klabukov, Denis Baranovskii, Viktoria Shishkina, Igor Buchwalow, Markus Tiemann and Dmitrii Atiakshin
Int. J. Mol. Sci. 2026, 27(7), 2923; https://doi.org/10.3390/ijms27072923 - 24 Mar 2026
Viewed by 733
Abstract
In idiopathic azoospermia caused by non-obstructive infertility with AZFc deletion, the testicle usually contains an increased number of mast cells (MCs)—which are responsible for collagen synthesis in the testes—as well as Leydig cell hyperplasia. However, the relationship between MCs and telocytes in this [...] Read more.
In idiopathic azoospermia caused by non-obstructive infertility with AZFc deletion, the testicle usually contains an increased number of mast cells (MCs)—which are responsible for collagen synthesis in the testes—as well as Leydig cell hyperplasia. However, the relationship between MCs and telocytes in this pathology remains unexplored. The aim of this study was to examine ultrastructural changes in the interstitial tissue microenvironment of the convoluted seminiferous tubules in the testis, using clinical specimens from men with genetically determined non-obstructive infertility with AZFc deletion. Histological, immunohistochemical, and electron microscopic (EM) studies were performed on surgical materials from 14 patients with AZFc deletion. The IHC study was performed using a panel of antibodies: tryptase, chymase, carboxypeptidase A3, and αSMA. The EM study was performed on ultrathin sections with a thickness of 100–120 nm. MCs were found to be in a functionally active state and characterized by a variety of secretory activities. For the first time, telocytes and their colocalization with MCs and Leydig cells were visualized. It is possibly the telocytes—interacting with MCs—that synchronize the functional activity of the entire MC population of the testis. The interaction of MCs with telocytes, as well as individual secretory granules associated with loci of tropocollagen and collagen microfibril accumulation, leads to the accumulation of collagen fibrils in the interstitium, as observed in idiopathic infertility with AZFc deletion. Even with a small number of MCs in the interstitium of the convoluted seminiferous tubules in the testis, the telocytes are able to synchronize MCs’ activation and secretory activity, supporting the development of a profibrotic phenotype of the tissue microenvironment. The obtained results advance our understanding of idiopathic infertility with AZFc deletion by delineating the ultrastructural landscape of the testicular interstitium and establishing telocytes as key regulators of cellular crosstalk. Telocytes use complex mechanisms for the spatial integration of MCs and fibroblasts in the profibrotic phenotype formation of the convoluted seminiferous tubule tissue microenvironment. Potentially, telocytes can directly be involved in synchronizing such processes by activating the biogenesis and secretion of collagen monomers by fibroblasts; the MC secretome directly affects the polymerization of collagen monomers and dimers into microfibrils in the extracellular matrix, stimulating excessive collagen fiber formation and the development of fibrotic changes. Full article
(This article belongs to the Special Issue Telocytes: Unraveling Their Roles in Health and Disease)
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18 pages, 2536 KB  
Article
The Allelopathic Inhibition of Submerged Macrophytes (Ceratophyllum demersum and Myriophyllum spicatum) in Response to Toxic and Non-Toxic Microcystis aeruginosa
by Yuanyuan Tang, Shuwen Zhang, Jing Dong, Yuanpu Sha, Guiyu Chen, Xuejun Li, Xiaofei Gao, Huatao Yuan, Jingxiao Zhang, Penghui Zhu and Yunni Gao
Microorganisms 2025, 13(12), 2797; https://doi.org/10.3390/microorganisms13122797 - 8 Dec 2025
Viewed by 1014
Abstract
The present study systematically explored the purification effects and response of submerged plants, Ceratophyllum demersum and Myriophyllum spicatum, on toxic and non-toxic strains of Microcystis aeruginosa via indoor co-culture experiments. The results showed that: (1) Both plants significantly inhibited the growth of [...] Read more.
The present study systematically explored the purification effects and response of submerged plants, Ceratophyllum demersum and Myriophyllum spicatum, on toxic and non-toxic strains of Microcystis aeruginosa via indoor co-culture experiments. The results showed that: (1) Both plants significantly inhibited the growth of Microcystis and reduced the concentration of chlorophyll-a (Chla) in the water by rapidly absorbing nutrients such as nitrogen and phosphorus, with no significant differences in the inhibition between toxic and non-toxic strains, indicating that nutrient competition might be the dominant mechanism for algal suppression. (2) C. demersum had higher nitrogen and phosphorus removal efficiency than M. spicatum, but the microcystins (MCs) released by toxic M. aeruginosa inhibited the nutrient removal capacity of both plants. (3) The plants promoted cell lysis of toxic M. aeruginosa and reduced extracellular MCs in the water while accumulating MCs internally, with C. demersum showing stronger MC accumulation and removal ability. (4) Microcystis stress activated the plants’ antioxidant defense systems, increased activities of SOD (Superoxide Dismutase) and CAT (Catalase), and caused membrane lipid peroxidation, increased content of MDA (Malondialdehyde), with toxic M. aeruginosa inducing stronger oxidative stress, and M. spicatum being more severely affected. (5) Plant species and algal toxicity jointly drove changes in the attached microbial community structure. The rhizosphere of M. spicatum specifically enriched Bdellovibrionota, suggesting a potential microbial predation pathway for algal suppression, while C. demersum was more associated with Bacillus and other microbes with allelopathic potential. In summary, C. demersum performed better in nutrient removal, toxin accumulation, and physiological tolerance. This study provides further theoretical support for using submerged plants to regulate cyanobacterial blooms and remediate eutrophic water bodies. Full article
(This article belongs to the Collection Biodegradation and Environmental Microbiomes)
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16 pages, 15008 KB  
Article
Mast Cells, Pancreatic Stellate Cells, and Telocytes in Chronic Pancreatitis: Ultrastructural Study
by Irina Chekmareva, Andrey Kostin, Oksana Paklina, Dmitry Kalinin, Dmitry Suraev, Nikolay Karnaukhov, Alexander Alekhnovich, Atim Emaimo John, Viktoria Shishkina, Igor Buchwalow, Markus Tiemann and Dmitrii Atiakshin
Int. J. Mol. Sci. 2025, 26(20), 10169; https://doi.org/10.3390/ijms262010169 - 19 Oct 2025
Cited by 1 | Viewed by 1550
Abstract
Pancreatic inflammation and subsequent fibrosis drive serious disease complications. However, the pathogenesis of this process and the mechanisms underlying excessive extracellular matrix (ECM) deposition remain poorly understood. Our aim was to study intercellular interactions and ultrastructural changes in mast cells, pancreatic stellate cells, [...] Read more.
Pancreatic inflammation and subsequent fibrosis drive serious disease complications. However, the pathogenesis of this process and the mechanisms underlying excessive extracellular matrix (ECM) deposition remain poorly understood. Our aim was to study intercellular interactions and ultrastructural changes in mast cells, pancreatic stellate cells, and telocytes, as well as in the extracellular matrix in various degrees of pancreatic fibrosis. Histological, immunohistochemical, and electron microscopic (EM) studies were performed on surgical materials from 17 patients. Mapping of fibrosis fields was performed on scanned images using the QuPath software v0.6.0. The IHC study was performed using a panel of antibodies: CD34, CD117, and SMA. Fluorescent IHC was performed using a panel of antibodies: CD34 and CD117. The EM study was performed on ultrathin sections with a thickness of 100–120 nm. The functional activity of mast cells (MCs) increased in pancreatic fibrosis. Most of the MCs were in a degranulation state, with the formation of intercellular contacts. The activation of pancreatic stellate cells (PaSCs), which underwent ultrastructural and functional changes in pancreatic fibrosis that developed as a result of chronic pancreatitis (CP), was noted. Multiple plasmolemma discontinuities, telopode shortenings, and nuclear fragmentations were observed among telocytes (TCs). The presence of MCs in the inflammatory infiltrate, as well as the destruction of TCs with the activation of exosomal transport, plays an important role in the pathogenesis of fibrosis in CP and provides a promising therapeutic target for the treatment of this pathology. Full article
(This article belongs to the Special Issue Roles of Mast Cells in Immune-Induced Diseases)
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15 pages, 1093 KB  
Article
Effectiveness of Ultrasound-Guided Peritendinous Injection Treatment with Low Molecular Weight Hyaluronic Acid in Patients with Supraspinatus Tendinopathy
by Francesco Agostini, Alessandro de Sire, Alessio Savina, Giovanni Iudicelli, Andrea Fisicaro, Giacomo Camponogara, Marco Narciso, Alessio Fricano, Marco Conti, Umile Giuseppe Longo, Valter Santilli, Antonio Ammendolia, Massimiliano Mangone and Marco Paoloni
J. Clin. Med. 2025, 14(17), 6291; https://doi.org/10.3390/jcm14176291 - 5 Sep 2025
Cited by 1 | Viewed by 2873
Abstract
Background/Objectives: Tendinopathies represent a prevalent musculoskeletal condition characterized by load-dependent pain, stiffness, weakness, and impaired functionality. Current treatment includes therapeutic exercise, physical modalities and injective therapy. Hyaluronic acid (HA) is a fundamental component of the extracellular matrix and plays a crucial role [...] Read more.
Background/Objectives: Tendinopathies represent a prevalent musculoskeletal condition characterized by load-dependent pain, stiffness, weakness, and impaired functionality. Current treatment includes therapeutic exercise, physical modalities and injective therapy. Hyaluronic acid (HA) is a fundamental component of the extracellular matrix and plays a crucial role in tissue hydration, elasticity, and lubrication. This study aims to evaluate the effectiveness of ultrasound-guided injections of HA in improving pain symptoms and functionality in patients with supraspinatus tendinopathy. Methods: Patients with a confirmed diagnosis of supraspinatus tendinopathy, verified through ultrasound imaging, were included in the study. Patients underwent 3 ultrasound guidance injections (1/week) of Sodium Hyaluronate (Hyalotend, 20 mg/2 mL). Patients were evaluated at the baseline, 1 month (T1), 3 months (T2), 6 months (T3), and 1 year (T4) after the first injection through the VAS, the QuickDASH and the SF-12. Results: Twenty-four patients were enrolled. As regards the VAS there was a statistically significant reduction (p < 0.01) of averages of values over time. The scores collected through the QuickDASH questionnaire have a statistically significant variation over time (p < 0.001). The values collected through the SF-12 Mental Component Summary (MCS) questionnaire show a highly statistically significant variation over time (p < 0.005). The values collected through the SF-12 Physical Component Summary (PCS) questionnaire show a statistically highly significant change over time (p < 0.001). Conclusions: Our results suggest that HA (Hyalotend) injections could represent a viable therapeutic option for patients with supraspinatus tendinopathy in the short, medium, and long term. Further studies with larger patient samples and a control group are needed to better investigate the effects and the modalities of administration of HA in tendinopathies. Full article
(This article belongs to the Special Issue Management of Ligaments and Tendons Injuries)
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30 pages, 58453 KB  
Article
Time- and Dose-Dependent Effects of Irradiation on Endothelial and Tumor Endothelial Cells: Transcriptional, Molecular, and Functional Changes Driving Activation In Vitro and In Vivo
by Iva Santek, Gregor Sersa and Bostjan Markelc
Cancers 2025, 17(17), 2842; https://doi.org/10.3390/cancers17172842 - 29 Aug 2025
Viewed by 2307
Abstract
Background: Irradiation (IR) targets cancer cells, but also the tumor microenvironment, including the tumor’s blood vessels. In addition to tumor endothelial cell (TEC) apoptosis, IR can lead to TEC activation, potentially increasing immune cell infiltration. However, the changes underlying the IR-induced activation of [...] Read more.
Background: Irradiation (IR) targets cancer cells, but also the tumor microenvironment, including the tumor’s blood vessels. In addition to tumor endothelial cell (TEC) apoptosis, IR can lead to TEC activation, potentially increasing immune cell infiltration. However, the changes underlying the IR-induced activation of endothelial cells (ECs) are poorly understood. This study investigated dose- and time-dependent molecular and functional responses of murine and human EC lines to IR in vitro and TECs in vivo in murine tumor models of colorectal carcinoma. Methods: HUVEC, EA.hy926, and Hulec5a, as well as murine bEND.3, 2H11, and SVEC4-10 EC lines, were irradiated with single doses of 2–10 Gy. EC proliferation and survival after IR were assessed by staining all nuclei (Hoechst 33342) and dead cells (propidium iodide) every 24 h for 5 days using the Cytation 1 Cell Imaging Multi-Mode Reader. RNA sequencing analysis of HUVECs irradiated with 2 Gy and 5 Gy at 24 h and 72 h after IR was conducted, focusing on processes related to EC activation. To validate the RNA sequencing results, immunofluorescence staining for proteins related to EC activation, including Stimulator of Interferon Response cGAMP Interactor 1 (STING), Nuclear factor kappa B (NF-κβ), and Vascular cell adhesion molecule 1 (VCAM-1), was performed. To validate the in vitro results, the response of TEC in vivo was analyzed using publicly available RNA sequencing data of TECs isolated from MC38 colon carcinoma irradiated with a single dose of 15 Gy. Finally, murine CT26 colon carcinoma tumors were immunofluorescently stained for STING and NF-κβ 24 and 48 h after IR with a clinically relevant fractionated regimen of 5 × 5 Gy. Results: Doses of 2, 4, 6, 8, and 10 Gy led to a dose-dependent decrease in proliferation and increased death of ECs. RNA sequencing analysis showed that the effects on the transcriptome of HUVECs were most pronounced 72 h after IR with 5 Gy, with 1014 genes (661 down-regulated and 353 up-regulated) being significantly differentially expressed. Irradiation with 5 Gy resulted in HUVEC activation, with up-regulation of the immune system and extracellular matrix genes, such as STING1 (log2FC = 0.81) and SELE (log2FC = 1.09), respectively; and down-regulation of cell cycle markers. Furthermore, IR led to the up-regulation of immune response- and extracellular matrix (ECM)-associated signaling pathways, including NF-κβ signaling and ECM–receptor interaction, which was also observed in the transcriptome of irradiated murine TECs in vivo. This was confirmed at the protein level with higher expressions of the EC activation-associated proteins STING, NF-κβ, and VCAM-1 in irradiated HUVECs and irradiated TECs in vivo. Conclusions: IR induces changes in ECs and TECs, supporting their activation in dose- and time-dependent manners, potentially contributing to the anti-tumor immune response, which may potentially increase the infiltration of immune cells into the tumor and thus, improve the overall efficacy of RT, especially in combination with immune checkpoint inhibitors. Full article
(This article belongs to the Special Issue Radiosensitivity and Radiotoxicity in Cancer)
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12 pages, 2819 KB  
Article
Dictamnus dasycarpus Turcz. Root Bark Improves Skin Barrier Function and Symptoms of Atopic Dermatitis in Mice
by Sangjun Park, Jinkyu Yang, Kyoungmin Sun, Seonah Park, Jimi Lee, Soyeon Kim, Ji Hyo Lyu and Hyungwoo Kim
Int. J. Mol. Sci. 2024, 25(23), 13178; https://doi.org/10.3390/ijms252313178 - 7 Dec 2024
Cited by 6 | Viewed by 2933
Abstract
The root bark of Dictamus dasycarpus Turcz. has been traditionally used for the topical treatment of skin disorders like pruritus. This study was designed to investigate the inflammatory and skin barrier protective effects of D. dasycarpus in mice with calcipotriol (MC903)-induced atopic dermatitis [...] Read more.
The root bark of Dictamus dasycarpus Turcz. has been traditionally used for the topical treatment of skin disorders like pruritus. This study was designed to investigate the inflammatory and skin barrier protective effects of D. dasycarpus in mice with calcipotriol (MC903)-induced atopic dermatitis (AD). Topical skin lesions on male Balb/c mice (8 weeks old) were treated topically with an ethanolic extract of D. dasycarpus (EEDD), and skin water content, water holding capacity (WHC), histopathological abnormalities, and inflammatory cytokine and chemokine levels were investigated. Topical application of EEDD effectively alleviated skin lesion severity, improved skin water content and WHC, and ameliorated histopathological abnormalities, including hyperkeratosis, blood vessel numbers near the epidermis, spongiotic changes, and immune cell infiltration in skin tissues. EEDD also suppressed inflammatory cytokines and chemokines, such as tumor necrosis factor (TNF)-α, thymic stromal lymphopoietin (TSLP), interleukin (IL)-1β, IL-4, IL-8, and monocyte chemotactic protein (MCP)-1. In RAW264.7 cells, EEDD reduced nuclear factor kappa-light-chain-enhancer of activated B cells (NF-kB) expression and suppressed the phosphorylations of extracellular signal-regulated kinase (ERK) and p38. These results suggest that the root bark of D. dasycarpus has therapeutic potential due to its anti-dermatitis and skin barrier protective effects in AD and that it could be used as an ingredient in skincare products. Full article
(This article belongs to the Section Bioactives and Nutraceuticals)
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14 pages, 2407 KB  
Article
Low, but Not High, Pulsating Fluid Shear Stress Affects Matrix Extracellular Phosphoglycoprotein Expression, Mainly via Integrin β Subunits in Pre-Osteoblasts
by Jianfeng Jin and Behrouz Zandieh-Doulabi
Curr. Issues Mol. Biol. 2024, 46(11), 12428-12441; https://doi.org/10.3390/cimb46110738 - 4 Nov 2024
Cited by 4 | Viewed by 1644
Abstract
Matrix extracellular phosphoglycoprotein (Mepe), present in bone and dentin, plays important multifunctional roles in cell signaling, bone mineralization, and phosphate homeostasis. Mepe expression in bone cells changes in response to pulsating fluid shear stress (PFSS), which is transmitted into cells through integrin-based adhesion [...] Read more.
Matrix extracellular phosphoglycoprotein (Mepe), present in bone and dentin, plays important multifunctional roles in cell signaling, bone mineralization, and phosphate homeostasis. Mepe expression in bone cells changes in response to pulsating fluid shear stress (PFSS), which is transmitted into cells through integrin-based adhesion sites, i.e., α and β subunits. Whether and to what extent PFSS influences Mepe expression through the modulation of integrin α and/or β subunit expression in pre-osteoblasts is uncertain. Therefore, we aimed to test whether low and/or high PFSS affects Mepe expression via modulation of integrin α and/or β subunit expression. MC3T3-E1 pre-osteoblasts were treated with ± 1 h PFSS (magnitude: 0.3 Pa (low PFSS) or 0.7 Pa (high PFSS); frequency: 1 Hz). Single integrin fluorescence intensity in pre-osteoblasts was increased, but single integrin area was decreased by low and high PFSS. Expression of two integrin α subunit-related genes (Itga1 and Itga5 2) was increased by low PFSS, and one (Itga5 2) by high PFSS. Expression of five integrin β subunit genes (Itgb1, Itgb3, Itgb5, Itgb5 13, and Itgb5 123) was increased by low PFSS, and three (Itgb5, Itgb5 13, and Itgb5 123) by high PFSS. Interestingly, Mepe expression in pre-osteoblasts was only modulated by low, but not high, PFSS. In conclusion, both low and high PFSS affected integrin α and β subunit expression in pre-osteoblasts, while integrin β subunit expression was more altered by low PFSS. Importantly, Mepe gene expression was only affected by low PFSS. These results might explain the different ways that Mepe-induced changes in pre-osteoblast mechanosensitivity may drive signaling pathways of bone cell function at low or high impact loading. These findings might have physiological and biomedical implications and require future research specifically addressing the precise role of integrin α or β subunits and Mepe during dynamic loading in bone health and disease. Full article
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22 pages, 18508 KB  
Article
The Role of Mast Cells in the Remodeling Effects of Molecular Hydrogen on the Lung Local Tissue Microenvironment under Simulated Pulmonary Hypertension
by Dmitrii Atiakshin, Andrey Kostin, Alexander Alekhnovich, Artem Volodkin, Michael Ignatyuk, Ilya Klabukov, Denis Baranovskii, Igor Buchwalow, Markus Tiemann, Marina Artemieva, Nataliya Medvedeva, Tyler W. LeBaron, Mami Noda and Oleg Medvedev
Int. J. Mol. Sci. 2024, 25(20), 11010; https://doi.org/10.3390/ijms252011010 - 13 Oct 2024
Cited by 5 | Viewed by 3930
Abstract
Molecular hydrogen (H2) has antioxidant, anti-inflammatory, and anti-fibrotic effects. In a rat model simulating pulmonary fibrotic changes induced by monocrotaline-induced pulmonary hypertension (MPH), we had previously explored the impact of inhaled H2 on lung inflammation and blood pressure. In this [...] Read more.
Molecular hydrogen (H2) has antioxidant, anti-inflammatory, and anti-fibrotic effects. In a rat model simulating pulmonary fibrotic changes induced by monocrotaline-induced pulmonary hypertension (MPH), we had previously explored the impact of inhaled H2 on lung inflammation and blood pressure. In this study, we further focused the biological effects of H2 on mast cells (MCs) and the parameters of the fibrotic phenotype of the local tissue microenvironment. MPH resulted in a significantly increased number of MCs in both the pneumatic and respiratory parts of the lungs, an increased number of tryptase-positive MCs with increased expression of TGF-β, activated interaction with immunocompetent cells (macrophages and plasma cells) and fibroblasts, and increased MC colocalization with a fibrous component of the extracellular matrix of connective tissue. The alteration in the properties of the MC population occurred together with intensified collagen fibrillogenesis and an increase in the integral volume of collagen and elastic fibers of the extracellular matrix of the pulmonary connective tissue. The exposure of H2 together with monocrotaline (MCT), despite individual differences between animals, tended to decrease the intrapulmonary MC population and the severity of the fibrotic phenotype of the local tissue microenvironment compared to changes in animals exposed to the MCT effect alone. In addition, the activity of collagen fibrillogenesis associated with MCs and the expression of TGF-β and tryptase in MCs decreased, accompanied by a reduction in the absolute and relative content of reticular and elastic fibers in the lung stroma. Thus, with MCT exposure, inhaled H2 has antifibrotic effects involving MCs in the lungs of rats. This reveals the unknown development mechanisms of the biological effects of H2 on the remodeling features of the extracellular matrix under inflammatory background conditions of the tissue microenvironment. Full article
(This article belongs to the Special Issue Physiology and Molecular Medicine of Molecular Hydrogen)
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13 pages, 2445 KB  
Article
A Critical Functional Missense Mutation (T117M) in Sheep MC4R Gene Significantly Leads to Gain-of-Function
by Ziyi Zhao, Yuta Yang, Peiyao Liu, Taotao Yan, Ran Li, Chuanying Pan, Yang Li and Xianyong Lan
Animals 2024, 14(15), 2207; https://doi.org/10.3390/ani14152207 - 30 Jul 2024
Cited by 3 | Viewed by 2002
Abstract
The melanocortin 4 receptor (MC4R) gene plays a central role in regulating energy homeostasis and food intake in livestock, thereby affecting their economic worth and growth. In a previous study, the p.T117M mutation in the sheep MC4R gene, which leads to [...] Read more.
The melanocortin 4 receptor (MC4R) gene plays a central role in regulating energy homeostasis and food intake in livestock, thereby affecting their economic worth and growth. In a previous study, the p.T117M mutation in the sheep MC4R gene, which leads to the transition of threonine to methionine, was found to affect the body weight at six months and the average daily gain in Hu sheep. However, there are still limited studies on the frequency of the sheep p.T117M missense mutation globally, and the underlying cellular mechanism remains elusive. Therefore, this study first used WGS to investigate the distribution of the MC4R gene p.T117M mutation in 652 individuals across 22 breeds worldwide. The results showed that the mutation frequency was higher in European breeds compared with Chinese sheep breeds, particularly in Poll Dorset sheep (mutation frequency > 0.5). The p.T117M mutation occurs in the first extracellular loop of MC4R. Mechanistically, the basal activity of the mutated receptor is significantly increased. Specifically, upon treatment with α-MSH and ACTH ligands, the cAMP and MAPK/ERK signaling activation of M117 MC4R is enhanced. These results indicate that the T117M mutation may change the function of the gene by increasing the constitutive activity and signaling activation of cAMP and MAPK/ERK, and, thus, may regulate the growth traits of sheep. In conclusion, this study delved into the global distribution and underlying cellular mechanisms of the T117M mutation of the MC4R gene, establishing a scientific foundation for breeding sheep with superior growth, thereby contributing to the advancement of the sheep industry. Full article
(This article belongs to the Section Animal Genetics and Genomics)
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12 pages, 247 KB  
Article
Evaluation of Change in Body Composition, including Phase Angle, in Post-Myocardial Infarction Patients Rehabilitated under the KOS-Zawał (MC-AMI) Programme
by Aleksandra Ślązak, Iga Przybylska and Małgorzata Paprocka-Borowicz
J. Clin. Med. 2024, 13(10), 2784; https://doi.org/10.3390/jcm13102784 - 9 May 2024
Cited by 6 | Viewed by 2452
Abstract
Background: Ischaemic heart disease, including myocardial infarction, is one of the main causes leading to heart failure as a consequence of ischaemic myocardial damage. In recent years, survival in the acute phase of myocardial infarction has improved significantly, but the high mortality rate [...] Read more.
Background: Ischaemic heart disease, including myocardial infarction, is one of the main causes leading to heart failure as a consequence of ischaemic myocardial damage. In recent years, survival in the acute phase of myocardial infarction has improved significantly, but the high mortality rate within 12 months of hospital discharge (reaching up to 9.8% in Poland) remains a challenge. Therefore, the KOS-Zawał (MC-AMI) comprehensive 12-month post-MI care programme was introduced in Poland in 2017. Aim: This study aimed to assess body composition (including, but not limited to, the phase angle, visceral fat, total body fat, redistribution between intracellular and extracellular fluid in the body, and metabolic age) using a bioelectrical impedance analysis (BIA) in post-MI patients before and after early post-MI rehabilitation who were participating in the KOS-Zawał (MC-AMI) programme. Methods: This study involved an examination (before rehabilitation) of 94 post-myocardial infarction patients who were referred to a cardiology appointment within 7–10 days of hospital discharge, during which a clinical assessment, electrocardiogram, and biochemical blood tests (complete blood count, CRP, and serum creatinine) were performed. For various reasons (death, qualification for device implantation, non-completion of rehabilitation, failure to attend a follow-up BIA), data from 55 patients who were examined twice (before and after rehabilitation) were used for the final analysis. Measurements were taken using a high-grade Tanita MC-780 BIA body composition analyser, which measured the resistance of tissues to a low-intensity electrical impulse (not perceptible to the subject). Results: Participation in rehabilitation as part of the KOS-Zawał (MC-AMI) programme was associated with a decrease in metabolic age in patients, with a reduction in visceral fat levels and levels of adipose tissue in the lower and upper limbs. Moreover, a clinically beneficial reduction in the ratio of extracellular water to total body water was also observed. These changes were statistically significant (p < 0.005). In contrast, there were no statistically significant differences in the change in phase angle values in the subjects before and after the 5-week post-infarction rehabilitation. Conclusions: Participation in early post-myocardial infarction rehabilitation as part of the KOS-Zawał (MC-AMI) programme (25 training sessions) is associated with significant improvements in body composition parameters, such as visceral adipose tissue, limb fat, and water redistribution, and, consequently, a reduction in metabolic age, despite no significant increase in phase angle values. It was hypothesised that the good baseline condition of the subjects might explain the lack of significant change in the phase angle over the short observation period. For further analysis, it would be worthwhile to increase the number of patients with baseline reduced phase angle values and monitor changes in this parameter throughout rehabilitation and the entire MC-AMI programme, because changes in the phase angle may also be influenced by other programme components such as dietary or psychological education. It is worth considering implementing a regular BIA assessment in patients in the programme as a motivating stimulus for diligent exercise and extending rehabilitation to be followed by telerehabilitation or hybrid telerehabilitation. Full article
(This article belongs to the Special Issue Recent Innovations in the Management of Cardiac Rehabilitation)
14 pages, 1600 KB  
Article
Changes in Extracellular Microcystins (MCs) Accompanying Algae/Cyanobacteria Removal during Three Representative Algae/Cyanobacteria Inactivation Processes and an MC Diffusion Model in Still Water
by Chengcheng Shi, Weijian Fang, Mengru Ma, Wei Xu and Jingjing Ye
Water 2023, 15(20), 3591; https://doi.org/10.3390/w15203591 - 13 Oct 2023
Cited by 4 | Viewed by 2711
Abstract
This study addresses the lack of comparative research on algae/cyanobacteria elimination technologies in the existing literature. Our investigation focused on evaluating the performance of three commonly used algae/cyanobacteria removal processes: ultrasound (20.8 kHz), copper sulfate and biotic algicide (Bacillus subtilis). The [...] Read more.
This study addresses the lack of comparative research on algae/cyanobacteria elimination technologies in the existing literature. Our investigation focused on evaluating the performance of three commonly used algae/cyanobacteria removal processes: ultrasound (20.8 kHz), copper sulfate and biotic algicide (Bacillus subtilis). The evaluation considered both algae/cyanobacteria removal efficacy and the consequent changes in extracellular microcystins (MCs). To achieve this, we employed real eutrophic water as the test water. The ultrasound treatment demonstrated effective algae/cyanobacteria removal, with an average rate of algae/cyanobacteria decreasing (RAD) ranging from 0.50 to 0.99 µg chlorophyll a per liter per minute (µg chlorophyll a/L·min). On the other hand, the copper sulfate and biotic algicide treatments exhibited relatively lower rates of algae/cyanobacteria removal, with average RAD values of 0.21 to 0.38 µg chlorophyll a per liter per day (µg chlorophyll a/L·d) and 0.10 to 0.13 µg chlorophyll a per liter per day (µg chlorophyll a/L·d), respectively. Moreover, we observed significant increases in extracellular MCs in the ultrasound and copper sulfate treatments. The corresponding values of the increment of extracellular MCs accompanying removal per microgramme (µg) chlorophyll a (IEMARMC) were 0.34 to 2.43 µg MCs per µg chlorophyll a (µg MCs/µg chlorophyll a) and 18.13 to 185.08 µg MCs per µg chlorophyll a (µg MCs/µg chlorophyll a), respectively. However, in certain conditions where sufficient dosages (0.5 to 2 mg/L) and reaction time (≥8 days) are provided, the biotic algicide treatment could result in a decrease in MCs compared to an untreated control group (IEMARMC: −43.94 to −32.18 µg MCs per µg chlorophyll a). This suggests that the biotic algicide effectively degraded the MCs. In addition, we developed a one-dimensional MC diffusion model in still water based on Newton’s second law, which exhibits excellent simulation capabilities. Full article
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15 pages, 10720 KB  
Article
Space-Flight- and Microgravity-Dependent Alteration of Mast Cell Population and Protease Expression in Digestive Organs of Mongolian Gerbils
by Dmitrii Atiakshin, Andrey Kostin, Viktoriya Shishkina, Alexandra Burtseva, Anastasia Buravleva, Artem Volodkin, Daniel Elieh-Ali-Komi, Igor Buchwalow and Markus Tiemann
Int. J. Mol. Sci. 2023, 24(17), 13604; https://doi.org/10.3390/ijms241713604 - 2 Sep 2023
Cited by 11 | Viewed by 2933
Abstract
Mast cell (MC)-specific proteases are of particular interest for space biology and medicine due to their biological activity in regulating targets of a specific tissue microenvironment. MC tryptase and chymase obtain the ability to remodel connective tissue through direct and indirect mechanisms. Yet, [...] Read more.
Mast cell (MC)-specific proteases are of particular interest for space biology and medicine due to their biological activity in regulating targets of a specific tissue microenvironment. MC tryptase and chymase obtain the ability to remodel connective tissue through direct and indirect mechanisms. Yet, MC-specific protease expression under space flight conditions has not been adequately investigated. Using immunohistochemical stainings, we analyzed in this study the protease profile of the jejunal, gastric, and hepatic MC populations in three groups of Mongolian gerbils—vivarium control, synchronous experiment, and 12-day orbital flight on the Foton-M3 spacecraft—and in two groups—vivarium control and anti-orthostatic suspension—included in the experiment simulating effects of weightlessness in the ground-based conditions. After a space flight, there was a decreased number of MCs in the studied organs combined with an increased proportion of chymase-positive MCs and MCs with a simultaneous content of tryptase and chymase; the secretion of specific proteases into the extracellular matrix increased. These changes in the expression of proteases were observed both in the mucosal and connective tissue MC subpopulations of the stomach and jejunum. Notably, the relative content of tryptase-positive MCs in the studied organs of the digestive system decreased. Space flight conditions simulated in the synchronous experiment caused no similar significant changes in the protease profile of MC populations. The space flight conditions resulted in an increased chymase expression combined with a decreased total number of protease-positive MCs, apparently due to participating in the processes of extracellular matrix remodeling and regulating the state of the cardiovascular system. Full article
(This article belongs to the Special Issue Cellular and Molecular Signaling Meet the Space Environment 2.0)
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