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37 pages, 3086 KiB  
Article
Conformal On-Body Antenna System Integrated with Deep Learning for Non-Invasive Breast Cancer Detection
by Marwa H. Sharaf, Manuel Arrebola, Khalid F. A. Hussein, Asmaa E. Farahat and Álvaro F. Vaquero
Sensors 2025, 25(15), 4670; https://doi.org/10.3390/s25154670 - 28 Jul 2025
Abstract
Breast cancer detection through non-invasive and accurate techniques remains a critical challenge in medical diagnostics. This study introduces a deep learning-based framework that leverages a microwave radar system equipped with an arc-shaped array of six antennas to estimate key tumor parameters, including position, [...] Read more.
Breast cancer detection through non-invasive and accurate techniques remains a critical challenge in medical diagnostics. This study introduces a deep learning-based framework that leverages a microwave radar system equipped with an arc-shaped array of six antennas to estimate key tumor parameters, including position, size, and depth. This research begins with the evolutionary design of an ultra-wideband octagram ring patch antenna optimized for enhanced tumor detection sensitivity in directional near-field coupling scenarios. The antenna is fabricated and experimentally evaluated, with its performance validated through S-parameter measurements, far-field radiation characterization, and efficiency analysis to ensure effective signal propagation and interaction with breast tissue. Specific Absorption Rate (SAR) distributions within breast tissues are comprehensively assessed, and power adjustment strategies are implemented to comply with electromagnetic exposure safety limits. The dataset for the deep learning model comprises simulated self and mutual S-parameters capturing tumor-induced variations over a broad frequency spectrum. A core innovation of this work is the development of the Attention-Based Feature Separation (ABFS) model, which dynamically identifies optimal frequency sub-bands and disentangles discriminative features tailored to each tumor parameter. A multi-branch neural network processes these features to achieve precise tumor localization and size estimation. Compared to conventional attention mechanisms, the proposed ABFS architecture demonstrates superior prediction accuracy and interpretability. The proposed approach achieves high estimation accuracy and computational efficiency in simulation studies, underscoring the promise of integrating deep learning with conformal microwave imaging for safe, effective, and non-invasive breast cancer detection. Full article
16 pages, 1113 KiB  
Case Report
Novel Sonoguided Digital Palpation and Ultrasound-Guided Hydrodissection of the Long Thoracic Nerve for Managing Serratus Anterior Muscle Pain Syndrome: A Case Report with Technical Details
by Nunung Nugroho, King Hei Stanley Lam, Theodore Tandiono, Teinny Suryadi, Anwar Suhaimi, Wahida Ratnawati, Daniel Chiung-Jui Su, Yonghyun Yoon and Keneath Dean Reeves
Diagnostics 2025, 15(15), 1891; https://doi.org/10.3390/diagnostics15151891 - 28 Jul 2025
Abstract
Background and Clinical Significance: Serratus Anterior Muscle Pain Syndrome (SAMPS) is an underdiagnosed cause of anterior chest wall pain, often attributed to myofascial trigger points of the serratus anterior muscle (SAM) or dysfunction of the Long Thoracic Nerve (LTN), leading to significant disability [...] Read more.
Background and Clinical Significance: Serratus Anterior Muscle Pain Syndrome (SAMPS) is an underdiagnosed cause of anterior chest wall pain, often attributed to myofascial trigger points of the serratus anterior muscle (SAM) or dysfunction of the Long Thoracic Nerve (LTN), leading to significant disability and affecting ipsilateral upper limb movement and quality of life. Current diagnosis relies on exclusion and physical examination, with limited treatment options beyond conservative approaches. This case report presents a novel approach to chronic SAMPS, successfully diagnosed using Sonoguided Digital Palpation (SDP) and treated with ultrasound-guided hydrodissection of the LTN using 5% dextrose in water (D5W) without local anesthetic (LA), in a patient where conventional treatments had failed. Case Presentation: A 72-year-old male presented with a three-year history of persistent left chest pain radiating to the upper back, exacerbated by activity and mimicking cardiac pain. His medical history included two percutaneous coronary interventions. Physical examination revealed tenderness along the anterior axillary line and a positive hyperirritable spot at the mid axillary line at the 5th rib level. SDP was used to visualize the serratus anterior fascia (SAF) and LTN, and to reproduce the patient’s concordant pain by palpating the LTN. Ultrasound-guided hydrodissection of the LTN was then performed using 20–30cc of D5W without LA to separate the nerve from the surrounding tissues, employing a “fascial unzipping” technique. The patient reported immediate pain relief post-procedure, with the pain reducing from 9/10 to 1/10 on the Numeric Rating Scale (NRS), and sustained relief and functional improvement at the 12-month follow-up. Conclusions: Sonoguided Digital Palpation (SDP) of the LTN can serve as a valuable diagnostic adjunct for visualizing and diagnosing SAMPS. Ultrasound-guided hydrodissection of the LTN with D5W without LA may provide a promising and safe treatment option for patients with chronic SAMPS refractory to conservative management, resulting in rapid and sustained pain relief. Further research, including controlled trials, is warranted to evaluate the long-term efficacy and generalizability of these findings and to compare D5W to other injectates. Full article
17 pages, 45942 KiB  
Article
Long-Term Culture of Cellular Spheroids in Novel Hydrogel Constructs for ECM Characterization in Bone Models
by Diamante Boscaro, Lill Skovholt Wahlum, Marie Eline Ullevålseter, Berit Løkensgard Strand and Pawel Sikorski
Materials 2025, 18(15), 3538; https://doi.org/10.3390/ma18153538 - 28 Jul 2025
Abstract
The application of cellular spheroids in bone tissue engineering research has gained significant interest in the last decade. Compared to monolayer cell cultures, the 3D architecture allows for more physiological cell–cell and cell–extracellular matrix (ECM) interactions that make cellular spheroids a suitable model [...] Read more.
The application of cellular spheroids in bone tissue engineering research has gained significant interest in the last decade. Compared to monolayer cell cultures, the 3D architecture allows for more physiological cell–cell and cell–extracellular matrix (ECM) interactions that make cellular spheroids a suitable model system to investigate the bone ECM in vitro. The use of 3D model systems requires fine-tuning of the experimental methods used to study cell morphology, ECM deposition and mineralization, and cell–ECM interactions. In this study, we use a construct made of MC3T3-E1 cellular spheroids encapsulated in an alginate hydrogel to study and characterize the deposited ECM. Spheroid shape and structure were evaluated using confocal microscopy. The deposited collagenous ECM was characterized using Second Harmonic Imaging Microscopy (SHIM), quantitative hydroxyproline (HYP) assay, and Transmission Electron Microscopy (TEM). The use of hydrogel constructs enabled easy handling and imaging of the samples, while also helping to preserve the spheroid’s stability by preventing cells from adhering to the culture dish surface. We used a non-modified alginate hydrogel that did not facilitate cell attachment and therefore functioned as an inert encapsulating scaffold. Constructs were cultured for up to 4 weeks. SHIM, HYP assay, and TEM confirmed the deposition of a collagenous matrix. We demonstrated that alginate-encapsulated bone spheroids are a convenient and promising model for studying the bone ECM in vitro. Full article
(This article belongs to the Section Biomaterials)
9 pages, 1013 KiB  
Article
Continuous Radiofrequency for Morton’s Neuroma: Is There Complete Ablation? A Preliminary Report
by Gabriel Camuñas-Nieves, Alejandro Fernández-Gibello, Simone Moroni, Felice Galluccio, Mario Fajardo-Pérez, Francisco Martínez-Pérez, Eduardo Simón-Pérez and Alfonso Martínez-Nova
Healthcare 2025, 13(15), 1838; https://doi.org/10.3390/healthcare13151838 - 28 Jul 2025
Abstract
Background and Objectives: Morton’s neuroma is a painful foot condition that can be treated with continuous radiofrequency. However, its efficacy is not always optimal, with failure rates of 15–20%. It has been suggested that these failures may be due to incomplete nerve ablation, [...] Read more.
Background and Objectives: Morton’s neuroma is a painful foot condition that can be treated with continuous radiofrequency. However, its efficacy is not always optimal, with failure rates of 15–20%. It has been suggested that these failures may be due to incomplete nerve ablation, allowing for nerve regeneration and persistent pain. So, the aim of this study was to assess the histological effects of continuous radiofrequency on the nerves affected by Morton’s neuroma. Materials and Methods: The effect of continuous radiofrequency was evaluated in two patients with Morton’s neuroma, which required open surgery excision. In both cases, radiofrequency with a standard protocol was applied ex vivo, following the surgical excision of the neuroma. A TLG10 RF generator (90 °C, 90 s) with a monopolar needle with a 0.5 cm active tip was used. Subsequently, the samples were histologically analyzed to determine the degree of nerve ablation. Results: Histological analysis showed homogeneous focal necrosis in both cases, with lesion depths of 2.4 mm and 3.18 mm. However, areas of intact nerve tissue were identified at the periphery of the neuroma, suggesting incomplete ablation. Conclusions: The findings indicate that continuous radiofrequency does not guarantee total nerve ablation, which could explain recurrence in some cases. Intraoperative neurophysiological monitoring could be key to optimizing the procedure, ensuring complete interruption of nerve conduction and improving treatment efficacy. Full article
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41 pages, 3039 KiB  
Review
Repurposing Diabetes Therapies in CKD: Mechanistic Insights, Clinical Outcomes and Safety of SGLT2i and GLP-1 RAs
by Syed Arman Rabbani, Mohamed El-Tanani, Rakesh Kumar, Manita Saini, Yahia El-Tanani, Shrestha Sharma, Alaa A. A. Aljabali, Eman Hajeer and Manfredi Rizzo
Pharmaceuticals 2025, 18(8), 1130; https://doi.org/10.3390/ph18081130 - 28 Jul 2025
Abstract
Background: Chronic Kidney Disease (CKD) is a major global health issue, with diabetes being its primary cause and cardiovascular disease contributing significantly to patient mortality. Recently, two classes of medications—sodium–glucose cotransporter 2 inhibitors (SGLT2i) and glucagon-like peptide-1 receptor agonists (GLP-1 RAs)—have shown promise [...] Read more.
Background: Chronic Kidney Disease (CKD) is a major global health issue, with diabetes being its primary cause and cardiovascular disease contributing significantly to patient mortality. Recently, two classes of medications—sodium–glucose cotransporter 2 inhibitors (SGLT2i) and glucagon-like peptide-1 receptor agonists (GLP-1 RAs)—have shown promise in protecting both kidney and heart health beyond their effects on blood sugar control. Methods: We conducted a narrative review summarizing the findings of different clinical trials and mechanistic studies evaluating the effect of SGLT2i and GLP-1 RAs on kidney function, cardiovascular outcomes, and overall disease progression in patients with CKD and DKD. Results: SGLT2i significantly mitigate kidney injury by restoring tubuloglomerular feedback, reducing intraglomerular hypertension, and attenuating inflammation, fibrosis, and oxidative stress. GLP-1 RAs complement these effects by enhancing endothelial function, promoting weight and blood pressure control, and exerting direct anti-inflammatory and anti-fibrotic actions on renal tissues. Landmark trials—CREDENCE, DAPA-CKD, and EMPA-KIDNEY—demonstrate that SGLT2i reduce the risk of kidney failure and renal or cardiovascular death by 25–40% in both diabetic and non-diabetic CKD populations. Likewise, trials such as LEADER, SUSTAIN, and AWARD-7 confirm that GLP-1 RAs slow renal function decline and improve cardiovascular outcomes. Early evidence suggests that using both drugs together may offer even greater benefits through multiple mechanisms. Conclusions: SGLT2i and GLP-1 RAs have redefined the therapeutic landscape of CKD by offering organ-protective benefits that extend beyond glycemic control. Whether used individually or in combination, these agents represent a paradigm shift toward integrated cardiorenal-metabolic care. A deeper understanding of their mechanisms and clinical utility in both diabetic and non-diabetic populations can inform evidence-based strategies to slow disease progression, reduce cardiovascular risk, and improve long-term patient outcomes in CKD. Full article
(This article belongs to the Special Issue New Development in Pharmacotherapy of Kidney Diseases)
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18 pages, 1359 KiB  
Article
Flavone C-Glycosides from Dianthus superbus L. Attenuate Metabolic Dysfunction-Associated Steatotic Liver Disease (MASLD) via Multi-Pathway Regulations
by Ming Chu, Yingying Tong, Lei Zhang, Yu Zhang, Jun Dang and Gang Li
Nutrients 2025, 17(15), 2456; https://doi.org/10.3390/nu17152456 - 28 Jul 2025
Abstract
Background: The metabolic dysfunction-associated steatotic liver disease (MASLD) represents an escalating global health concern, with effective treatments still lacking. Given its complex pathogenesis, multi-targeted strategies are highly desirable. Methods: This study reports the isolation of four flavone C-glycosides (FCGs) from Dianthus superbus L. [...] Read more.
Background: The metabolic dysfunction-associated steatotic liver disease (MASLD) represents an escalating global health concern, with effective treatments still lacking. Given its complex pathogenesis, multi-targeted strategies are highly desirable. Methods: This study reports the isolation of four flavone C-glycosides (FCGs) from Dianthus superbus L. and explores their potential in treating MASLD. The bioactivity and underlying mechanisms of FCGs were systematically evaluated by integrating network pharmacology, molecular docking, and zebrafish model validation. Results: Network pharmacology analysis revealed that FCGs may modulate multiple MASLD-related pathways, including lipid metabolism, insulin signaling, inflammation, and apoptosis. Molecular docking further confirmed strong binding affinities between FCGs and key protein targets involved in these pathways. In the zebrafish model of MASLD induced by egg yolk powder, FCGs administration markedly attenuated obesity, hepatic lipid accumulation, and liver tissue damage. Furthermore, FCGs improved lipid metabolism and restored locomotor function. Molecular analyses confirmed that FCGs upregulated PPARγ expression to promote lipid metabolism, restored insulin signaling by enhancing INSR, PI3K, and AKT expression, and suppressed inflammation by downregulating TNF, IL-6 and NF-κB. Additionally, FCGs inhibited hepatocyte apoptosis by elevating the BCL-2/BAX ratio. Conclusions: These findings highlight the multi-pathway regulatory effects of FCGs in MASLD, underscoring its potential as a novel therapeutic candidate for further preclinical development. Full article
24 pages, 1413 KiB  
Article
Potential of Pumpkin Pulp Carotenoid Extract in the Prevention of Doxorubicin-Induced Cardiotoxicity
by Milana Bosanac, Alena Stupar, Biljana Cvetković, Dejan Miljković, Milenko Čanković and Bojana Andrejić Višnjić
Pharmaceutics 2025, 17(8), 977; https://doi.org/10.3390/pharmaceutics17080977 - 28 Jul 2025
Abstract
Background/Objectives: Doxorubicin is a chemotherapeutic agent whose clinical use is limited by side effects (SEs). The most common SE is doxorubicin-induced cardiotoxicity (DIC), for which there is still no prevention. The hypothesis arises that active substances of natural origin could influence DIC [...] Read more.
Background/Objectives: Doxorubicin is a chemotherapeutic agent whose clinical use is limited by side effects (SEs). The most common SE is doxorubicin-induced cardiotoxicity (DIC), for which there is still no prevention. The hypothesis arises that active substances of natural origin could influence DIC prevention by affecting several pathways of DIC occurrence. Methods: Thirty Wistar rats were divided into six groups (control, NADES (C8:C10) solvent, pumpkin pulp extract, doxorubicin, NADES (C8:C10) solvent–doxorubicin, and pumpkin pulp extract–doxorubicin). During the experiment, parameters of general condition, body, and heart weight were observed. Heart function parameters were monitored by measuring the levels of serum NT-pro-BNP, CK-MB, and hsTnT. Tissue damage was evaluated by determining the doxorubicin damage score and the expression of anti-cardiac troponin I, anti-Nrf2, anti-Bcl-2, anti-caspase-3, anti-COX2, and anti-Ki67 antibodies. Results: Doxorubicin administration led to impaired general condition of animals and increased the levels of NT-proBNP, CK-MB, hsTnT, and myocardium tissue damage of medium grade. Its administration induced apoptosis (as evidenced by elevated Casp3), reduced antiapoptotic Bcl-2 and troponin I expression in cardiomyocytes. Reduced Nrf2 expression due to doxorubicin administration was restored when pumpkin pulp extract containing carotenoids was coadministered, which led to the normalization of Casp3, Bcl-2, and troponin I expression. Consequently, the general condition and body weight were better in animals treated with both doxorubicin and the other treatment compared to those treated with doxorubicin alone. Conclusions: The results of this study strongly suggest that pumpkin pulp extract containing carotenoids has a cardioprotective effect, possibly by regulating the Nrf2 pathway. Full article
(This article belongs to the Special Issue Plant Extracts and Their Biomedical Applications)
17 pages, 2045 KiB  
Article
Different Phosphorus Preferences Among Arbuscular and Ectomycorrhizal Trees with Different Acquisition Strategies in a Subtropical Forest
by Yaping Zhu, Jianhua Lv, Pifeng Lei, Miao Chen and Jinjuan Xie
Forests 2025, 16(8), 1241; https://doi.org/10.3390/f16081241 - 28 Jul 2025
Abstract
Phosphorus (P) availability is a major constraint on plant growth in many forest ecosystems, yet the strategies by which different tree species acquire and utilize various forms of soil phosphorus remain poorly understood. This study investigated how coexisting tree species with contrasting mycorrhizal [...] Read more.
Phosphorus (P) availability is a major constraint on plant growth in many forest ecosystems, yet the strategies by which different tree species acquire and utilize various forms of soil phosphorus remain poorly understood. This study investigated how coexisting tree species with contrasting mycorrhizal types, specifically arbuscular mycorrhizal (AM) and ectomycorrhizal (ECM) associations, respond to different phosphorus forms under field conditions. An in situ root bag experiment was conducted using four phosphorus treatments (control, inorganic, organic, and mixed phosphorus) across four subtropical tree species. A comprehensive set of fine root traits, including morphological, physiological, and mycorrhizal characteristics, was measured to evaluate species-specific phosphorus foraging strategies. The results showed that AM species were more responsive to phosphorus form variation than ECM species, particularly under inorganic and mixed phosphorus treatments. Significant changes in root diameter (RD), root tissue density (RTD), and acid phosphatase activity (RAP) were observed in AM species, often accompanied by higher phosphorus accumulation in fine roots. For example, RD in AM species significantly decreased under the Na3PO4 treatment (0.94 mm) compared to the control (1.18 mm), while ECM species showed no significant changes in RD across treatments (1.12–1.18 mm, p > 0.05). RTD in AM species significantly increased under Na3PO4 (0.030 g/cm3) and Mixture (0.021 g/cm3) compared to the control (0.012 g/cm3, p < 0.05), whereas ECM species exhibited consistently low RTD values across treatments (0.017–0.020 g/cm3, p > 0.05). RAP in AM species increased significantly under Na3PO4 (1812 nmol/g/h) and Mixture (1596 nmol/g/h) relative to the control (1348 nmol/g/h), while ECM species showed limited variation (1286–1550 nmol/g/h, p > 0.05). In contrast, ECM species displayed limited trait variation across treatments, reflecting a more conservative acquisition strategy. In addition, trait correlation analysis revealed stronger coordination among root traits in AM species. And AM species exhibited high variability across treatments, while ECM species maintained consistent trait distributions with limited plasticity. These findings suggest that AM and ECM species adopt fundamentally different phosphorus acquisition strategies. AM species rely on integrated morphological and physiological responses to variable phosphorus conditions, while ECM species maintain stable trait configurations, potentially supported by fungal symbiosis. Such divergence may contribute to functional complementarity and species coexistence in phosphorus-limited subtropical forests. Full article
(This article belongs to the Section Forest Ecophysiology and Biology)
26 pages, 1183 KiB  
Review
The Art of Interpreting Antinuclear Antibodies (ANAs) in Everyday Practice
by Marcelina Kądziela, Aleksandra Fijałkowska, Marzena Kraska-Gacka and Anna Woźniacka
J. Clin. Med. 2025, 14(15), 5322; https://doi.org/10.3390/jcm14155322 - 28 Jul 2025
Abstract
Background: Antinuclear antibodies (ANAs) serve as crucial biomarkers for diagnosing systemic autoimmune diseases; however, their interpretation can be complex and may not always correlate with clinical symptoms. Methods: A comprehensive narrative review was conducted to evaluate the peer-reviewed literature published between 1961 and [...] Read more.
Background: Antinuclear antibodies (ANAs) serve as crucial biomarkers for diagnosing systemic autoimmune diseases; however, their interpretation can be complex and may not always correlate with clinical symptoms. Methods: A comprehensive narrative review was conducted to evaluate the peer-reviewed literature published between 1961 and 2025. Databases, including PubMed and Scopus, were searched using combinations of controlled vocabulary and free-text terms relating to antinuclear antibodies and their clinical significance. The objective was to gather and synthesize information regarding the diagnostic utility and interpretation of ANA testing in routine medical practice. Discussion: The indirect immunofluorescence assay (IIF) on HEp-2 cells is established as the gold standard for detecting ANAs, facilitating the classification of various fluorescent patterns. While a positive ANA test can suggest autoimmune disorders, the presence and titre must be interpreted alongside clinical findings, as low titres often lack diagnostic significance. Findings indicate that titres higher than 1:160 may provide greater specificity in differentiating true positives from false positives in healthy individuals. The study also emphasizes the relevance of fluorescence patterns, with specific patterns linked to particular diseases, although many do not have strong clinical correlations. Moreover, certain autoantibodies demonstrate high specificity for diseases like systemic lupus erythematosus (SLE) and mixed connective tissue disease (MCTD). Ultimately, while ANA testing is invaluable for diagnosing connective tissue diseases, healthcare providers must consider its limitations to avoid misdiagnosis and unnecessary treatment. Conclusions: ANA testing is a valuable tool in the diagnosis of connective tissue diseases, but its interpretation must be approached with caution. Clinical context remains crucial when evaluating ANA results to avoid misdiagnosis and overtreatment. This review is about the diagnostic aspects and clinical consequences of ANA testing, as well as highlighting both the diagnostic benefits and the potential limitations of this procedure in everyday clinical practice. The review fills a gap in the literature by integrating the diagnostic and clinical aspects of ANA testing, with a focus on real-world interpretation challenges. Full article
(This article belongs to the Section Immunology)
16 pages, 3457 KiB  
Article
Association Between Genotoxic Effects of Ageing Dental Restorations and Micronuclei in Oral Mucosal Cells
by Csilla Benedek, Bernadette Kerekes-Máthé, Zsuzsanna Bardocz-Veres, Boglárka Szabó, Alina Iacob, Alexandra Stoica, Timea Dako, Mónika Kovács, Lóránd Dénes and Liana Bereșescu
Medicina 2025, 61(8), 1363; https://doi.org/10.3390/medicina61081363 - 28 Jul 2025
Abstract
Background and Objectives: Dental restorations can be composed of various materials, including amalgams and methacrylate-based resins. The health risks associated with the components of the restorative materials have always been a concern, even more so with the ageing of the restorations. As [...] Read more.
Background and Objectives: Dental restorations can be composed of various materials, including amalgams and methacrylate-based resins. The health risks associated with the components of the restorative materials have always been a concern, even more so with the ageing of the restorations. As the micronucleus (MN) test is a standard, accessible, and minimally invasive technique for studying the genotoxic effect of clastogenic chemicals on oral mucosal cells, the current study was conducted to determine the frequency and morphological properties of MN in the exfoliated oral mucosal cells. Materials and Methods: A total of 115 aged composite and amalgam restorations were included in this study. Epithelial cells were collected from the gingival tissue adjacent to the restorations of each patient and stained with a hematoxylin–eosin (HE) stain. After evaluation of the slides, the results were subjected to statistical analysis using Chi-square tests. The level of significance was set at 0.05. Results: The mean number of MN was significantly lower for composite restorations compared to amalgam restorations. There were no statistically significant differences between composite restorations aged 1–5 years, 5–10 years, over 10 years, and amalgam restorations aged over 10 years in the location (p = 0.11), staining (p = 0.11), or morphological characteristics (p = 0.18) of the MN. Conclusions: Despite the main limitation of this study, the lack of a control group, our results suggest that long-term exposure to restorative fillings and the ageing of these materials can cause DNA damage locally in the adjacent sites of oral cavity. Full article
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22 pages, 6477 KiB  
Article
Human Small Intestinal Tissue Models to Assess Barrier Permeability: Comparative Analysis of Caco-2 Cells, Jejunal and Duodenal Enteroid-Derived Cells, and EpiIntestinalTM Tissues in Membrane-Based Cultures with and Without Flow
by Haley L. Moyer, Leoncio Vergara, Clifford Stephan, Courtney Sakolish, Hsing-Chieh Lin, Weihsueh A. Chiu, Remi Villenave, Philip Hewitt, Stephen S. Ferguson and Ivan Rusyn
Bioengineering 2025, 12(8), 809; https://doi.org/10.3390/bioengineering12080809 - 28 Jul 2025
Abstract
Accurate in vitro models of intestinal permeability are essential for predicting oral drug absorption. Standard models like Caco-2 cells have well-known limitations, including lack of segment-specific physiology, but are widely used. Emerging models such as organoid-derived monolayers and microphysiological systems (MPS) offer enhanced [...] Read more.
Accurate in vitro models of intestinal permeability are essential for predicting oral drug absorption. Standard models like Caco-2 cells have well-known limitations, including lack of segment-specific physiology, but are widely used. Emerging models such as organoid-derived monolayers and microphysiological systems (MPS) offer enhanced physiological relevance but require comparative validation. We performed a head-to-head evaluation of Caco-2 cells, human jejunal (J2) and duodenal (D109) enteroid-derived cells, and EpiIntestinalTM tissues cultured on either static Transwell and flow-based MPS platforms. We assessed tissue morphology, barrier function (TEER, dextran leakage), and permeability of three model small molecules (caffeine, propranolol, and indomethacin), integrating the data into a physiologically based gut absorption model (PECAT) to predict human oral bioavailability. J2 and D109 cells demonstrated more physiologically relevant morphology and higher TEER than Caco-2 cells, while the EpiIntestinalTM model exhibited thicker and more uneven tissue structures with lower TEER and higher passive permeability. MPS cultures offered modest improvements in epithelial architecture but introduced greater variability, especially with enteroid-derived cells. Predictions of human fraction absorbed (Fabs) were most accurate when using static Caco-2 data with segment-specific corrections based on enteroid-derived values, highlighting the utility of combining traditional and advanced in vitro gut models to optimize predictive performance for Fabs. While MPS and enteroid-based systems provide physiological advantages, standard static models remain robust and predictive when used with in silico modeling. Our findings support the need for further refinement of enteroid-MPS integration and advocate for standardized benchmarking across gut model systems to improve translational relevance in drug development and regulatory reviews. Full article
(This article belongs to the Section Biomedical Engineering and Biomaterials)
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24 pages, 4347 KiB  
Article
Diatomaceous Earth Supplementation as a Bioavailable Silicon Source Modulates Postprandial Lipid Metabolism in Healthy Female Rats
by Rocío Redondo-Castillejo, Marina Hernández-Martín, Jousef Ángel Issa-García, Aránzazu Bocanegra, Alba Garcimartín, Adrián Macho-González, Sara Bastida, Francisco J. Sánchez-Muniz, Juana Benedí and M. Elvira López-Oliva
Nutrients 2025, 17(15), 2452; https://doi.org/10.3390/nu17152452 - 28 Jul 2025
Abstract
Background/Objectives: Diatomaceous earth (DE), a natural substance rich in amorphous silica and recognized as a food additive, is gaining attention as a dietary silicon supplement. However, its bioavailability and impact on lipid digestion and absorption remain poorly characterized. This study aimed to investigate [...] Read more.
Background/Objectives: Diatomaceous earth (DE), a natural substance rich in amorphous silica and recognized as a food additive, is gaining attention as a dietary silicon supplement. However, its bioavailability and impact on lipid digestion and absorption remain poorly characterized. This study aimed to investigate silicon bioavailability after short-term DE supplementation and its effects on postprandial glycemia and triglyceridemia, the expression of lipid metabolism-related proteins, and the modulation of the intestinal mucosal barrier. Methods: Female Wistar rats received daily oral supplementation of DE (equivalent to 2 or 4 mg silicon/kg body weight) for one week. Silicon digestibility, excretion, and hepatic accumulation were quantified. Postprandial glycemia and triglyceridemia were monitored. Lipid profile was analyzed by HPSEC in gastric and intestinal contents. Jejunal morphology and mucin-secreting cells were assessed histologically. Lipid metabolism markers were evaluated by immunohistochemistry and Western blot in both intestinal and hepatic tissues. Results: DE supplementation enhanced silicon absorption and increased hepatic levels. Fecal output and moisture content were also elevated, especially at the higher dose. DE significantly reduced postprandial triglyceridemia and consequently increased luminal triglyceride retention. These changes were associated with decreased jejunal levels of IFABP, ACAT2, and MTP, as well as reduced hepatic levels of MTP and LDLr, alongside increased levels of ABCG5/G8 and LXRα/β, indicating a partial blockage of lipid absorption and enhanced cholesterol efflux. The effects on the intestinal barrier were evidenced by villi shortening and an increase in mucin-producing cells. Conclusion: Food-grade DE is a bioavailable source of silicon with hypolipidemic potential, mainly by reducing intestinal lipid absorption. This is supported by lower postprandial triglycerides, increased luminal lipid retention, and decreased expression of lipid transport proteins. The study in healthy female rats underscores the importance of sex-specific responses and supports DE as a dietary strategy to improve lipid metabolism. Full article
(This article belongs to the Section Lipids)
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21 pages, 1019 KiB  
Review
Macrophage Reprogramming: Emerging Molecular Therapeutic Strategies for Nephrolithiasis
by Meng Shu, Yiying Jia, Shuwei Zhang, Bangyu Zou, Zhaoxin Ying, Xu Gao, Ziyu Fang and Xiaofeng Gao
Biomolecules 2025, 15(8), 1090; https://doi.org/10.3390/biom15081090 - 28 Jul 2025
Abstract
Nephrolithiasis, predominantly driven by calcium oxalate (CaOx) crystal deposition, poses a significant global health burden due to its high prevalence and recurrence rates and limited preventive/therapeutic options. Recent research has underscored a pivotal role for macrophage polarization in nephrolithiasis pathogenesis. Pro-inflammatory phenotype macrophages [...] Read more.
Nephrolithiasis, predominantly driven by calcium oxalate (CaOx) crystal deposition, poses a significant global health burden due to its high prevalence and recurrence rates and limited preventive/therapeutic options. Recent research has underscored a pivotal role for macrophage polarization in nephrolithiasis pathogenesis. Pro-inflammatory phenotype macrophages exacerbate crystal-induced injury and foster stone formation by amplifying crystal adhesion via an NF-κB–IL-1β positive-feedback axis that sustains ROS generation and NLRP3 inflammasome activation, whereas anti-inflammatory phenotype macrophages facilitate crystal clearance and tissue repair. We have summarized the research on treating nephrolithiasis and related renal injury by targeting macrophage polarization in recent years, including therapeutic approaches through pharmacological methods, epigenetic regulation, and advanced biomaterials. At the same time, we have critically evaluated the novel therapeutic strategies for macrophage reprogramming and explored the future development directions of targeting macrophage reprogramming for nephrolithiasis treatment, such as using single-cell/spatial omics to reveal the heterogeneity of macrophages in the stone microenvironment, chimeric antigen receptor macrophages (CAR-Ms) as a potential therapy for specific crystal phagocytosis in certain areas, and multi-omics integration to address inter-patient immune differences. This review highlights that macrophage reprogramming is a transformative frontier in nephrolithiasis management and underscores the need for further research to translate these molecular insights into effective clinical applications. Full article
(This article belongs to the Section Molecular Medicine)
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27 pages, 1010 KiB  
Review
The Multifaceted Role of IL-35 in Periodontal Disease and Beyond: From Genetic Polymorphisms to Biomarker Potential
by Zdravka Pashova-Tasseva, Antoaneta Mlachkova, Kamen Kotsilkov and Hristina Maynalovska
Genes 2025, 16(8), 891; https://doi.org/10.3390/genes16080891 - 28 Jul 2025
Abstract
Periodontitis is a prevalent chronic inflammatory disease with complex etiopathogenesis involving microbial dysbiosis, host immune response, environmental factors, and genetic susceptibility. Among the cytokines implicated in periodontal immunoregulation, interleukin-35 (IL-35) has emerged as a novel anti-inflammatory mediator with potential diagnostic and therapeutic relevance. [...] Read more.
Periodontitis is a prevalent chronic inflammatory disease with complex etiopathogenesis involving microbial dysbiosis, host immune response, environmental factors, and genetic susceptibility. Among the cytokines implicated in periodontal immunoregulation, interleukin-35 (IL-35) has emerged as a novel anti-inflammatory mediator with potential diagnostic and therapeutic relevance. This narrative review evaluates the role of IL-35 in periodontal disease by exploring its local and systemic expression, response to non-surgical periodontal therapy (NSPT), and association with clinical disease severity. Additionally, current evidence regarding IL-35 gene polymorphisms and their potential contribution to individual susceptibility and disease progression, as well as their relevance in related systemic conditions, is assessed. A comprehensive review and synthesis of recent clinical and experimental studies were conducted, focusing on IL-35 levels in saliva, serum, and gingival crevicular fluid (GCF) among patients with healthy periodontium, gingivitis, and various stages of periodontitis, both before and after NSPT. Emphasis was placed on longitudinal studies evaluating IL-35 dynamics in correlation with periodontal parameters, as well as genetic association studies investigating IL-12A and EBI3 gene polymorphisms. IL-35 levels were generally found to be higher in healthy individuals and reduced in periodontitis patients, indicating a possible protective role in maintaining periodontal homeostasis. Following NSPT, IL-35 levels significantly increased, corresponding with clinical improvement and reduced inflammatory burden. Genetic studies revealed variable associations between IL-35 polymorphisms and susceptibility to periodontitis and related systemic conditions, although further research is needed for validation. IL-35 appears to function as a modulator of immune resolution in periodontal disease, with potential utility as a non-invasive biomarker for disease activity and therapeutic response. Its upregulation during periodontal healing supports its role in promoting tissue stabilization. The integration of cytokine profiling and genetic screening may enhance personalized risk assessment and targeted interventions in periodontal care. Full article
(This article belongs to the Section Human Genomics and Genetic Diseases)
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27 pages, 4829 KiB  
Article
Quantitative Analysis of Ginger Maturity and Pulsed Electric Field Thresholds: Effects on Microstructure and Juice’s Nutritional Profile
by Zhong Han, Pan He, Yu-Huan Geng, Muhammad Faisal Manzoor, Xin-An Zeng, Suqlain Hassan and Muhammad Talha Afraz
Foods 2025, 14(15), 2637; https://doi.org/10.3390/foods14152637 - 28 Jul 2025
Abstract
This study used fresh (young) and old (mature) ginger tissues as model systems to investigate how plant maturity modulates the response to pulsed electric field (PEF), a non-thermal processing technology. Specifically, the influence of tissue maturity on dielectric behavior and its downstream effect [...] Read more.
This study used fresh (young) and old (mature) ginger tissues as model systems to investigate how plant maturity modulates the response to pulsed electric field (PEF), a non-thermal processing technology. Specifically, the influence of tissue maturity on dielectric behavior and its downstream effect on juice yield and bioactive compound extraction was systematically evaluated. At 2.5 kV/cm, old ginger exhibited a pronounced dielectric breakdown effect due to enhanced electrolyte content and cell wall lignification, resulting in a higher degree of cell disintegration (0.65) compared with fresh ginger (0.44). This translated into a significantly improved juice yield of 90.85% for old ginger, surpassing the 84.16% limit observed in fresh ginger. HPLC analysis revealed that the extraction efficiency of 6-gingerol and 6-shogaol increased from 1739.16 to 2233.60 µg/g and 310.31 to 339.63 µg/g, respectively, in old ginger after PEF treatment, while fresh ginger showed increases from 1257.88 to 1824.05 µg/g and 166.43 to 213.52 µg/g, respectively. Total phenolic content (TPC) and total flavonoid content (TFC) also increased in both tissues, with OG-2.5 reaching 789.57 µg GAE/mL and 336.49 µg RE/mL, compared with 738.19 µg GAE/mL and 329.62 µg RE/mL in FG-2.5. Antioxidant capacity, as measured by ABTS•+ and DPPH inhibition, improved more markedly in OG-2.5 (37.8% and 18.7%, respectively) than in FG-2.5. Moreover, volatile compound concentrations increased by 177.9% in OG-2.5 and 137.0% in FG-2.5 compared with their respective controls, indicating differential aroma intensification and compound transformation. Structural characterization by SEM and FT-IR further corroborated enhanced cellular disruption and biochemical release in mature tissue. Collectively, these results reveal a maturity-dependent mechanism of electro-permeabilization in plant tissues, offering new insights into optimizing non-thermal processing for functional food production. Full article
(This article belongs to the Section Food Engineering and Technology)
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