Biomedicines

18 pages, 2646 KiB

Open AccessArticle

The IL-6/JAK/STAT3 Axis in Cholangiocarcinoma and Primary Sclerosing Cholangitis: Unlocking Therapeutic Strategies Through Patient-Derived Organoids

by Corinna Boden, Laura K. Esser, Leona Dold, Bettina Langhans, Taotao Zhou, Dominik J. Kaczmarek, Maria A. Gonzalez-Carmona, Tobias J. Weismüller, Glen Kristiansen, Jörg C. Kalff, Michael Hölzel, Hanno Matthaei, Marieta I. Toma and Vittorio Branchi

Biomedicines 2025, 13(5), 1083; https://doi.org/10.3390/biomedicines13051083 (registering DOI) - 29 Apr 2025

Abstract

Background/Objectives: Primary sclerosing cholangitis (PSC) is a rare, incurable liver disease characterized by chronic biliary inflammation and fibrosis. PSC is a significant risk factor for biliary tract cancer (BTC). This study aims to evaluate STAT3 expression in BTC and its prognostic significance as [...] Read more.

Background/Objectives: Primary sclerosing cholangitis (PSC) is a rare, incurable liver disease characterized by chronic biliary inflammation and fibrosis. PSC is a significant risk factor for biliary tract cancer (BTC). This study aims to evaluate STAT3 expression in BTC and its prognostic significance as well as explore the potential of organoids derived from PSC and liver tumor patients as an in vitro model for testing novel therapeutic strategies in both PSC and BTC. Methods: Fresh tissue samples obtained from 10 PSC patients through targeted endoscopic retrograde cholangiography (ERC) and biopsy samples from liver tumor patients were used to establish organoid cultures. Organoids were treated with different agents and the therapeutic effect was measured by CellTiterGlo. Treatment with the JAK inhibitor baricitinib was followed by the measurement of cytokine concentrations in the supernatant. Archived formalin-fixed paraffin-embedded (FFPE) samples from 55 surgically resected BTC tumors were analyzed for STAT3 expression using immunohistochemistry. Results: We successfully established organoid cultures from all ERC samples. STAT3 protein expression was detected in 56% of tumor samples and 69% of the immune microenvironment. STAT3 positivity in the immune cell compartment was associated with longer disease-free survival, although the multivariate analysis could not confirm its value as an independent prognostic factor. Chemotherapy testing on liver tumor organoids showed various degrees of decreases in viability after treatment with gemcitabine, cisplatin, and cabozantinib. Baricitinib treatment significantly reduced IL-6 and MCP-1 secretion in cholangiocarcinoma Conclusions: The patient-derived organoid model of PSC and liver tumors is a valuable tool for testing novel and established therapeutic strategies, including JAK inhibitors and chemotherapy regimens. STAT3 expression in the immune microenvironment of BTC may serve as a prognostic marker. Further studies are needed to explore the integration of co-cultured organoid systems with stromal and immune components to improve physiological relevance. Full article

(This article belongs to the Special Issue Recent Advances in Gastrointestinal Cancers: From Microbiota Modulation to New Therapeutic Approaches)

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21 pages, 635 KiB

Open AccessReview

The Emerging Role and Mechanism of E2/E3 Hybrid Enzyme UBE2O in Human Diseases

by Qian Cheng, Zuyin Li, Yongjian Li, Lei Chen, Dingbao Chen and Jiye Zhu

Biomedicines 2025, 13(5), 1082; https://doi.org/10.3390/biomedicines13051082 (registering DOI) - 29 Apr 2025

Abstract

The ubiquitin–proteasome system (UPS) plays a pivotal role in determining protein fate, regulating signal transduction, and maintaining cellular homeostasis. Protein ubiquitination, a key post-translational modification, is orchestrated by the sequential actions of three primary enzymes, ubiquitin-activating enzyme (E1), ubiquitin-conjugating enzyme (E2), and ubiquitin [...] Read more.

The ubiquitin–proteasome system (UPS) plays a pivotal role in determining protein fate, regulating signal transduction, and maintaining cellular homeostasis. Protein ubiquitination, a key post-translational modification, is orchestrated by the sequential actions of three primary enzymes, ubiquitin-activating enzyme (E1), ubiquitin-conjugating enzyme (E2), and ubiquitin protein ligase (E3), alongside the regulatory influence of deubiquitinases (DUBs) and various cofactors. The process begins with E1, which activates ubiquitin molecules. Subsequently, E2 receives the activated ubiquitin from E1 and transfers it to E3. E3, in turn, recognizes specific target proteins and facilitates the covalent attachment of ubiquitin from E2 to lysine residues on the target protein. Among the E2 enzymes, ubiquitin-conjugating enzyme E2O (UBE2O) stands out as a unique E2–E3 hybrid enzyme. UBE2O directly mediates the ubiquitination of a wide array of substrates, including 5′-AMP-activated protein kinase catalytic subunit alpha-2 (AMPKα2), MAX interactor 1 (Mxi1), and v-maf musculoaponeurotic fibrosarcoma oncogene homolog (c-Maf), among others. In this narrative review, we will explore the structural characteristics of UBE2O and elucidate its molecular functions. Additionally, we will summarize recent advancements in understanding the role of UBE2O in various tumors, Alzheimer’s disease (AD), and metabolic diseases. Finally, we will discuss the potential of targeting UBE2O as a novel therapeutic strategy for the treatment of human diseases. Full article

(This article belongs to the Special Issue Ubiquitylation and Deubiquitylation in Health and Diseases)

28 pages, 1250 KiB

Open AccessReview

Advancing Antidepressive Agents: Drug Discovery and Polymer-Based Drug Delivery Systems for Improved Treatment Outcome

by Yufei Zhang, Zengyi Song, Hongxi Zhang, Haijiao Lin, Pu Xu, Zijia Li, Qingyun He and Binbin Wei

Biomedicines 2025, 13(5), 1081; https://doi.org/10.3390/biomedicines13051081 - 29 Apr 2025

Abstract

Depressive disorder (a subclass of mental disorders) is characterized by persistent affective symptoms. Without timely therapeutic intervention, it leads to clinical deterioration manifested as reduced quality of life and may increase suicide risk in severe cases. Given its complex etiology, intertwined with intrinsic [...] Read more.

Depressive disorder (a subclass of mental disorders) is characterized by persistent affective symptoms. Without timely therapeutic intervention, it leads to clinical deterioration manifested as reduced quality of life and may increase suicide risk in severe cases. Given its complex etiology, intertwined with intrinsic factors such as genetics and environment, and impacted by various issues such as first-pass effect and blood-brain barrier, the therapeutic efficacy of many antidepressant medications is limited for patients. Therefore, by delving into the exploration of novel antidepressant drugs and biomaterials, this review aims to offer fresh perspectives that may facilitate the discovery of innovative antidepressant medications and enhance their therapeutic outcomes. Notably, the review highlights polymers’ crucial role in enhancing antidepressants’ pharmacological efficacy and pharmacokinetic properties by optimizing their parameters, and they will undoubtedly become powerful tools in improving antidepressive outcomes in future research. Full article

(This article belongs to the Section Neurobiology and Clinical Neuroscience)

18 pages, 2660 KiB

Open AccessArticle

Developing CGMap: Characterizing Continuous Glucose Monitoring Data in Patients with Type 2 Diabetes

by Shuzhen Bai, Chu Lin, Xiaoling Cai, Suiyuan Hu, Jing Wu, Ling Chen, Wenjia Yang and Linong Ji

Biomedicines 2025, 13(5), 1080; https://doi.org/10.3390/biomedicines13051080 - 29 Apr 2025

Abstract

Objectives: This study will characterize continuous glucose monitoring (CGM) data in patients with type 2 diabetes in China, and assess the relationship between CGM-derived indicators and diabetes-related clinical parameters. Methods: The data for this study were collected from a randomized trial [...] Read more.

Objectives: This study will characterize continuous glucose monitoring (CGM) data in patients with type 2 diabetes in China, and assess the relationship between CGM-derived indicators and diabetes-related clinical parameters. Methods: The data for this study were collected from a randomized trial in China (ChiCTR2000039424) from February 2020 to July 2022 in which patients wore a CGM device for 14 days. Glycemia risk index (GRI), coefficient of variation (CV), standard deviation (SD), mean amplitude of glycemic excursions (MAGE), time in range (TIR), time above range (TAR), time below range (TBR), and estimate glycated hemoglobin (eA1c) were analyzed. Ordinary least square linear regression and the Spearman method were used to test the relationship between CGM-derived indicators and diabetes-related clinical parameters. Results: In all, 528 patients with type 2 diabetes from a randomized controlled trial were analyzed. It was shown that CV, SD, and MAGE increased with age and diabetes duration, but decreased with an increase in body mass index. Higher fasting plasma glucose, higher baseline HbA1c, and higher insulin resistance levels were associated with higher GRI, SD, MAGE, TAR, and eA1c, and they were associated with lower TIR. In addition, higher HOMA-2β was associated with higher TIR and TBR, and with lower TAR and eA1c. Hemoglobin had positive correlations to SD, TAR, and eA1c. Conclusions: It was found that glucose variability increased with age and the duration of diabetes. However, glucose variability decreased with increased BMI. Meanwhile, greater glycemic variability was associated with worse islet function, higher baseline glucose level, and higher hemoglobin. Full article

(This article belongs to the Section Endocrinology and Metabolism Research)

13 pages, 1534 KiB

Open AccessArticle

The Effects of the Biological Agents Infliximab, Vedolizumab, and Ustekinumab on Intestinal Anastomosis: An Experimental Study in Rats

by Alexandra Menni, Georgios Tzikos, Patroklos Goulas, George Chatziantoniou, Angeliki Vouchara, Athanasios S. Apostolidis, Aristeidis Ioannidis, Georgios Germanidis, Lyssimachos G. Papazoglou, Olga Giouleme and Stylianos Apostolidis

Biomedicines 2025, 13(5), 1079; https://doi.org/10.3390/biomedicines13051079 - 29 Apr 2025

Abstract

Background/Objectives: The potential side effects of the use of biological agents in the perioperative period are still under investigation. This animal prospective study aimed to evaluate the overall impact of biological factor administration after intestinal surgery. Methods: This study included 80 [...] Read more.

Background/Objectives: The potential side effects of the use of biological agents in the perioperative period are still under investigation. This animal prospective study aimed to evaluate the overall impact of biological factor administration after intestinal surgery. Methods: This study included 80 female Wistar rats sorted into four groups: three groups received one of the biological factors, infliximab, vedolizumab, or ustekinumab; the control group received placebo therapy. After enterotomy and intestinal anastomosis, the bursting pressure (BP) of the anastomosis was compared among the groups on postoperative days (PODs) 3 and 7. Results: On POD3, the control group presented with a significantly higher mean BP (154.6 ± 39.7 mmHg) compared to the infliximab (66.8 ± 10.4 mmHg), vedolizumab (105.4 ± 37.6 mmHg), and ustekinumab (98.8 ± 47.9 mmHg) groups. A post hoc analysis among the three biological agent groups revealed differences only when comparing infliximab and vedolizumab rats with the controls on POD3 (p < 0.001) and with the ustekinumab rats on POD7, having a greater mean BP (282.5 ± 80.1 mmHg, p = 0.031). No differences were observed regarding the event of broken anastomosis among the four groups. Conclusions: This experimental study’s findings highlight the varying detrimental effects of different biological agents on the strength of intestinal anastomosis, with ustekinumab demonstrating superior performance. Full article

(This article belongs to the Special Issue Pancreatic, Liver, Biliary Tract and Intestinal Diseases: Pathogenesis, Diagnostics and Therapy)

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11 pages, 13128 KiB

Open AccessCase Report

Posterior Mitral Valve Hypoplasia: Three Clinical Cases and a Review of the Literature

by Baudouin Koenig, Alin Ionescu and Elena Galli

Biomedicines 2025, 13(5), 1078; https://doi.org/10.3390/biomedicines13051078 - 29 Apr 2025

Abstract

Background: Posterior mitral leaflet hypoplasia is a rare condition, generally diagnosed in children. This paper presents three adult cases of posterior leaflet hypoplasia and comprehensively reviews the existing literature. Methods and Results: All patients presented with severe mitral regurgitation necessitating hospitalization. Transthoracic and [...] Read more.

Background: Posterior mitral leaflet hypoplasia is a rare condition, generally diagnosed in children. This paper presents three adult cases of posterior leaflet hypoplasia and comprehensively reviews the existing literature. Methods and Results: All patients presented with severe mitral regurgitation necessitating hospitalization. Transthoracic and transesophageal echocardiography were performed to determine the underlying etiology, revealing pronounced posterior mitral leaflet hypoplasia. Each case was evaluated by a multidisciplinary heart team. Surgical mitral valve intervention was feasible in two patients. In one high-risk patient, percutaneous treatment options for MR were explored but ultimately deemed unsuitable due to the anatomical characteristics of the valve. The patient was consequently managed conservatively with medical therapy. Conclusions: These cases demonstrate that posterior mitral leaflet hypoplasia may constitute an underrecognized cause of severe primary mitral regurgitation in adults. The management of such cases can be particularly challenging due to the atypical valvular morphology. Full article

(This article belongs to the Section Molecular and Translational Medicine)

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16 pages, 4839 KiB

Open AccessArticle

Differential Effects of Canonical Androgens and 11-Ketotestosterone on Reproductive Phenotypes and Folliculogenesis in Mouse Model of PCOS

by Yi-Ru Tsai, Yen-Nung Liao, Cheng-Ju Tsai, Yu-Ang Lee, Shih-Min Hsia, Kuo-Chung Lan and Hong-Yo Kang

Biomedicines 2025, 13(5), 1077; https://doi.org/10.3390/biomedicines13051077 - 29 Apr 2025

Abstract

Background: Polycystic ovary syndrome (PCOS) is a common female endocrine disorder characterized by hyperandrogenism, ovulatory dysfunction, and polycystic ovarian morphology. While canonical androgens like testosterone (T) and dihydrotestosterone (DHT) are well studied in PCOS pathophysiology, the role of 11-ketotestosterone (11KT) remains unclear. [...] Read more.

Background: Polycystic ovary syndrome (PCOS) is a common female endocrine disorder characterized by hyperandrogenism, ovulatory dysfunction, and polycystic ovarian morphology. While canonical androgens like testosterone (T) and dihydrotestosterone (DHT) are well studied in PCOS pathophysiology, the role of 11-ketotestosterone (11KT) remains unclear. This study investigates the differential effects of these androgens on folliculogenesis, ovulation, and steroidogenesis using in vivo and in vitro models. Methods: Four-week-old female C57BL/6 mice received T, DHT, or 11KT for six weeks. The assessments included body weight, estrous cyclicity, serum hormone profiles, ovarian histology, and follicle classification. In parallel, large preantral follicles were cultured with each androgen to evaluate follicle growth, antrum formation, and ovulation capacity. Androgen receptor (AR) signaling and steroidogenic function were analyzed using western blotting, RT-qPCR, and luciferase reporter assays. Results: The DHT-treated mice exhibited increased weight gain, whereas 11KT-treated mice showed reduced weight gain. T and DHT disrupted the estrous cycle, while 11KT prolonged diestrus. All androgen treatments led to ovarian morphological changes, including follicular arrest and cystic features. In vitro, all androgens enhanced follicle growth, but only T and DHT inhibited ovulation. The AR expression was elevated across all androgen-treated groups, but only DHT significantly activated AR and CYP19A1 promoters. Conclusions: 11KT induces a distinct and milder PCOS-like phenotype compared to classical androgens, promoting follicle growth with minimal impact on ovulation or steroidogenic disruption. These findings underscore the heterogeneity of PCOS and suggest that different androgen profiles may drive diverse clinical phenotypes. By elucidating the distinct roles of different androgens, this may lead to better stratification of PCOS phenotypes based on predominant androgen types for more precise diagnosis and individualized management. Full article

(This article belongs to the Special Issue Animal Models of Human Pathology: Revision, Relevance and Refinements (3rd Edition))

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17 pages, 1564 KiB

Open AccessReview

Diabetic Foot Ulcers: Pathophysiology, Immune Dysregulation, and Emerging Therapeutic Strategies

by John Dawi, Kevin Tumanyan, Kirakos Tomas, Yura Misakyan, Areg Gargaloyan, Edgar Gonzalez, Mary Hammi, Serly Tomas and Vishwanath Venketaraman

Biomedicines 2025, 13(5), 1076; https://doi.org/10.3390/biomedicines13051076 - 29 Apr 2025

Abstract

Diabetic foot ulcers (DFUs) are among the most common and debilitating complications of diabetes mellitus (DM), affecting approximately 15–25% of patients and contributing to over 85% of non-traumatic amputations. DFUs impose a substantial clinical and economic burden due to high recurrence rates, prolonged [...] Read more.

Diabetic foot ulcers (DFUs) are among the most common and debilitating complications of diabetes mellitus (DM), affecting approximately 15–25% of patients and contributing to over 85% of non-traumatic amputations. DFUs impose a substantial clinical and economic burden due to high recurrence rates, prolonged wound care, and frequent hospitalizations, accounting for billions in healthcare costs worldwide. The multifactorial pathophysiology of DFUs involves peripheral neuropathy, peripheral arterial disease, chronic inflammation, and impaired tissue regeneration. Recent studies underscore the importance of immune dysregulation—specifically macrophage polarization imbalance, regulatory T cell dysfunction, and neutrophil impairment—as central mechanisms in wound chronicity. These immune disruptions sustain a pro-inflammatory environment dominated by cytokines, such as TNF-α, IL-1β, and IL-6, which impair angiogenesis and delay repair. This review provides an updated synthesis of DFU pathogenesis, emphasizing immune dysfunction and its therapeutic implications. We examine emerging strategies in immunomodulation, regenerative medicine, and AI-based wound technologies, including SGLT2 inhibitors, biologics, stem cell therapies, and smart dressing systems. These approaches hold promise for accelerating healing, reducing amputation risk, and personalizing future DFU care. Full article

(This article belongs to the Special Issue New Diagnostic and Therapeutic Approaches in Diabetic Microvascular Complications, 2nd Edition)

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12 pages, 828 KiB

Open AccessArticle

Artificial Intelligence Algorithm to Screen for Diabetic Neuropathy: A Pilot Study

by Giovanni Sartore, Eugenio Ragazzi, Francesco Pegoraro, Mario German Pagno, Annunziata Lapolla and Francesco Piarulli

Biomedicines 2025, 13(5), 1075; https://doi.org/10.3390/biomedicines13051075 - 29 Apr 2025

Abstract

Background/Objectives: Patients with type 2 diabetes (T2D) are at risk of developing multiple complications, and diabetic polyneuropathy (DPN) is by far the most common. The purpose of the present study was to assess the ability of a new algorithm based on artificial [...] Read more.

Background/Objectives: Patients with type 2 diabetes (T2D) are at risk of developing multiple complications, and diabetic polyneuropathy (DPN) is by far the most common. The purpose of the present study was to assess the ability of a new algorithm based on artificial intelligence (AI) to identify patients with T2D who are at risk of DPN in order to move on to further instrumental evaluation with the biothesiometer method. Methods: This is a single-centre, cross-sectional study with 201 consecutive T2D patients recruited at the Diabetes Operating Unit of the ULSS 6 of Padua (Northeast Italy). The individual risk of developing DPN was calculated using the AI-based MetaClinic Prediction Algorithm and compared with the DPN diagnosis provided by the digital biothesiometer method, which measures the vibratory perception threshold (VPT) on both feet. Results: Of the enrolled patients, 107 (53.23%) were classified by AI software as having a low probability of developing DPN, 39 (19.40%) as having a moderate probability, 29 (14.43%) as having a high probability, and 26 (12.94%) as having a very high probability. In 63 of the total patients, biothesiometer measurement showed a VPT ≥ 25 V, indicative of DPN, while 138 patients had a non-pathological VPT value (< 25 V) (prevalence of abnormal VPT 31.34%; prevalence of normal VPT 68.66%). The overall agreement between biothesiometer results and AI risk attribution was 65%. Cohen’s κ was 0.162, and Gwet’s AC1 coefficient 0.405. Conclusions: The use of an optimized AI algorithm can help estimate the risk of developing DPN, thereby guiding more targeted and in-depth screening, including instrumental assessment using the biothesiometer method. Full article

(This article belongs to the Special Issue Diabetes: Comorbidities, Therapeutics and Insights (2nd Edition))

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Graphical abstract

12 pages, 1732 KiB

Open AccessArticle

Investigation of the Effect of Amniomax on Lidocaine-Induced Toxicity in Healthy Colon Cell Culture

by Seçil Azime Karakuş, Ayten Saraçoğlu, Eray Metin Güler, Kübra Bozali, Ceren Önal, Yekbun Bulun, Tomasz Gaszyński, Paweł Ratajczyk and Kemal Tolga Saraçoğlu

Biomedicines 2025, 13(5), 1074; https://doi.org/10.3390/biomedicines13051074 - 29 Apr 2025

Abstract

Background: Lidocaine (LIDO) toxicity is a critical concern in regional anesthesia, with no specific antidote currently available. While lipid emulsions are commonly used as rescue agents in cases of local anesthetic systemic toxicity (LAST), their efficacy is inconsistent, and their safety remains controversial. [...] Read more.

Background: Lidocaine (LIDO) toxicity is a critical concern in regional anesthesia, with no specific antidote currently available. While lipid emulsions are commonly used as rescue agents in cases of local anesthetic systemic toxicity (LAST), their efficacy is inconsistent, and their safety remains controversial. AmnioMax^® (AMX), a specialized cell culture medium enriched with growth factors and bioactive molecules, has the potential to offer cytoprotective effects. This study aims to investigate the therapeutic efficacy of AMX in mitigating lidocaine-induced cytotoxicity and to explore its protective mechanisms at the cellular level. Methods: Healthy colon cells (CCD-18Co) were used in this study. Four experimental groups were established as follows: control, LIDO, AMX, and LIDO + AMX. Cellular viability in the control group was set at 100%. LIDO was administered at concentrations ranging from 0.06 to 10%, AMX at 0.625–100%, and LIDO + AMX at 60% of the half-maximal effective concentration (EC₅₀) combined with LIDO (0.06–10%). Cells were incubated for 24 h, after which cellular viability, DNA damage, apoptosis, intracellular reactive oxygen species (iROS), intracellular calcium (Ca), mitochondrial membrane potential (MMP), and glutathione (GSH) were evaluated. Results: LIDO exposure led to a concentration-dependent decrease in viability compared to the control group (p < 0.001), while AMX significantly increased viability (p < 0.001). In the LIDO + AMX group, viability was also reduced (p < 0.001); however, cytotoxicity was significantly lower than in the LIDO group (p < 0.05). Both the LIDO and LIDO + AMX groups showed increased iROS levels, DNA damage, and apoptosis (p < 0.001), along with the decreased MMP and GSH levels (p < 0.001) compared to the control. However, in the LIDO + AMX group, iROS, DNA damage, and apoptosis were significantly lower than in the LIDO group (p < 0.01), MMP levels were increased (p < 0.001), and no significant difference was observed in GSH levels. Conclusions: AMX demonstrated cytoprotective effects against LIDO-induced cytotoxicity, suggesting its potential as an alternative therapeutic agent for LAST. Full article

(This article belongs to the Section Drug Discovery, Development and Delivery)

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11 pages, 1984 KiB

Open AccessArticle

Limited Performance of Machine Learning Models Developed Based on Demographic and Laboratory Data Obtained Before Primary Treatment to Predict Coronary Aneurysms

by Mi-Jin Kim, Gi-Beom Kim, Dongha Yang, Yeon-Jin Jang and Jeong-Jin Yu

Biomedicines 2025, 13(5), 1073; https://doi.org/10.3390/biomedicines13051073 - 29 Apr 2025

Abstract

Background/objectives: Kawasaki disease is the leading cause of acquired heart disease in children within developed countries. Although treatment with intravenous immunoglobulin (IVIG) significantly reduces the incidence of coronary artery aneurysm (CAA), the risk of it persists, affecting long-term patient outcomes. While intensified [...] Read more.

Background/objectives: Kawasaki disease is the leading cause of acquired heart disease in children within developed countries. Although treatment with intravenous immunoglobulin (IVIG) significantly reduces the incidence of coronary artery aneurysm (CAA), the risk of it persists, affecting long-term patient outcomes. While intensified primary treatment is recommended for patients at high risk of IVIG resistance or CAA development, a universally accepted predictive model for such resistance remains unestablished. This study aims to develop a machine learning model to predict the occurrence of CAAs prior to initiating IVIG therapy. Methods: Data from two nationwide epidemiological surveys conducted between 2012 and 2017 were analyzed, encompassing 17,189 patients with calculable coronary artery z-scores and Harada scores. Various supervised machine learning algorithms were applied to develop a model for predicting CAA. Afterward, unsupervised learning techniques were employed to explore the data’s inherent structure. Results: The Harada score’s receiver operating characteristic (ROC) analysis yielded an area under the curve (AUC) of 0.558. The highest AUC among the machine learning models was 0.661, achieved by the Light Gradient Boosting Machine. However, this model’s sensitivity was 0.615, and specificity was 0.647, indicating limited clinical applicability. Unsupervised learning revealed no distinct distribution patterns between patients with/without CAAs. Conclusions: Despite utilizing a large dataset to develop a machine learning-based prediction model for CAAs, the performance was unsatisfactory. Future studies should focus on enhancing predictive models by incorporating additional clinical data, such as acute-phase coronary artery diameter measurements, to improve accuracy and clinical utility. Full article

(This article belongs to the Special Issue Updates on Kawasaki Disease)

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11 pages, 1631 KiB

Open AccessArticle

SpO₂/FiO₂ Correlates with PaO₂/FiO₂ (P/F) and Radiological Biomarkers of Severity: A Retrospective Study on COVID-19 Pneumonia Patients

by Alberto Marra, Vito D’Agnano, Raffaella Pagliaro, Fabio Perrotta, Ilaria Di Fiore, Antonio D’Orologio, Filippo Scialò, Angela Schiattarella, Andrea Bianco and Roberto Parrella

Biomedicines 2025, 13(5), 1072; https://doi.org/10.3390/biomedicines13051072 - 28 Apr 2025

Abstract

Background: In patients with COVID-19 pneumonia, the estimation of PaO₂ represents the method of choice for monitoring a patient’s oxygenation status and assessing disease severity. The aim of this study is, therefore, to investigate the correlation between SpO₂/FiO₂ and [...] Read more.

Background: In patients with COVID-19 pneumonia, the estimation of PaO₂ represents the method of choice for monitoring a patient’s oxygenation status and assessing disease severity. The aim of this study is, therefore, to investigate the correlation between SpO₂/FiO₂ and PaO₂/FiO₂, as well as radiological and laboratory biomarkers of severity. Methods: In this monocentric observational, analytical, retrospective large cohort study, consecutive patients with a confirmed diagnosis of pneumonia from SARS-CoV-2, hospitalized at the Cotugno Hospital—AORN dei Colli—of Naples, between 1 September 2020 and 28 February 2022 were considered for study inclusion. Patients with missing data were excluded. Results: We included 585 patients (median age 63 [22–95]). Mean PaO₂/FiO₂ was 203 [66–433], whilst mean SpO₂/FiO₂ was 240 [81–471]. We found that P/F ratio could be predicted from S/F ratio, as described by the linear regression equation (P/F = 13.273 + 0.790 × S/F). In addition, we found that SpO₂/FiO₂ ratio significantly correlated with HRCT score and laboratory markers of severity, including IL-6, D-Dimer, and NLR. Conclusions: SpO₂/FiO₂ ratio represents a highly useful resource as a valid surrogate of P/F ratio in patients with COVID pneumonia, also correlating with other biomarkers of severity, such as HRCT score and key laboratory markers. Full article

(This article belongs to the Special Issue Advances in Lung Cancer: From Bench to Bedside)

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20 pages, 858 KiB

Open AccessArticle

PTX3/NF-κB/TLR4 Pathway Evaluation in the Follicular Fluid to Successfully Predict Blastocyst Implantation: A Pilot Study

by Alessio Ardizzone, Carmelo Liuzzo, Arianna Ferro, Marco Galletta, Emanuela Esposito and Anna Paola Capra

Biomedicines 2025, 13(5), 1071; https://doi.org/10.3390/biomedicines13051071 - 28 Apr 2025

Abstract

Background: The implantation process is complex and involves numerous factors that can affect its success. In artificial reproductive treatments (ARTs), chronic inflammation seems to be associated with implantation failure, largely contributing to reproductive dysfunction. Pentraxin 3 (PTX3) is overexpressed in several pathological conditions [...] Read more.

Background: The implantation process is complex and involves numerous factors that can affect its success. In artificial reproductive treatments (ARTs), chronic inflammation seems to be associated with implantation failure, largely contributing to reproductive dysfunction. Pentraxin 3 (PTX3) is overexpressed in several pathological conditions by exerting a pivotal role both as a regulator and indicator of inflammatory response. Some literature data have shown that PTX3 could have an impact on follicle growth and development, influencing women’s fertility. This study aimed to detect PTX3 in follicular fluids collected during an ART protocol in relation to implantation outcomes. Methods: The PTX3/NF-kB/TLR4 pathway and other cytokines were assessed in the follicular fluid of 169 subjects, under the age of 40 years, undergoing IVF cycles, including females without achieved implantation (n = 98) and those with implantation (n = 71). Furthermore, subgroup analyses were performed to evaluate PTX3 values according to age difference. Results: From our data, PTX3 emerged as a strong predictor, more than TNFα and IL-1β, of implantation failure and related inflammatory follicular state. Overall, the results point to PTX3 as a potential biomarker for ART success, and their testing may be helpful in women whose successful implantation remains unexplained. Conclusions: Therefore, PTX3 could constitute a reliable biomarker and a valuable target to improve ART outcomes. Full article

(This article belongs to the Special Issue Role of Factors in Embryo Implantation and Placental Development)

14 pages, 6715 KiB

Open AccessArticle

“Anti-Bios”: Can Local Antibiotics Affect Bone Union in Infected Bone Defects Treated with Degradable Bone Substitutes

by Filippo Vandenbulcke, Salvatore Lorenzo Renne, Giuseppe Anzillotti, Pietro Conte, Giuliano Ravasio, Gabriele Meroni, Federica Riva and Elizaveta Kon

Biomedicines 2025, 13(5), 1070; https://doi.org/10.3390/biomedicines13051070 - 28 Apr 2025

Abstract

Background: Segmental bone defects (SBDs) pose significant clinical challenges, often requiring complex reconstructive procedures. Degradable bone substitutes loaded with antibiotics have emerged as promising tools for infection control. However, their impact on bone healing remains uncertain. This study investigates antibiotic-loaded biodegradable scaffolds [...] Read more.

Background: Segmental bone defects (SBDs) pose significant clinical challenges, often requiring complex reconstructive procedures. Degradable bone substitutes loaded with antibiotics have emerged as promising tools for infection control. However, their impact on bone healing remains uncertain. This study investigates antibiotic-loaded biodegradable scaffolds in infected defects using an in vivo rabbit model. Methods: Thirty New Zealand white rabbits were divided into three groups—antibiotic-loaded GreenBone scaffolds, non-loaded GreenBone scaffolds, and allografts. A critical-size femoral defect was surgically created and inoculated with Staphylococcus epidermidis. Radiographic evaluations were performed over 16 weeks, followed by histological and microbiological analyses. Bone union, infection rates, and callus maturation were assessed. Results: Eight rabbits were excluded for technical errors. Bone union was significantly lower in the antibiotic-loaded group (two rabbits out of seven; 28.6%) compared to the non-loaded scaffold (13 rabbits out of 15; 86.7%; p = 0.006). The antibiotic-loaded group exhibited a higher incidence of chronic osteomyelitis (100%) versus non-loaded implants (60%; p < 0.05). Histological evaluation revealed delayed bone maturation in the antibiotic-loaded group (22.2% HOES grade 3) compared to non-loaded scaffolds (69.5%; p < 0.001). Conclusions: Despite their infection-fighting potential, antibiotic-loaded biodegradable scaffolds may impair bone healing, leading to higher non-union rates and delayed maturation. These findings highlight a critical trade-off between local antibiotic therapy and bone regeneration, warranting careful clinical consideration and further research to optimize treatment strategies for infected bone defects. Full article

(This article belongs to the Special Issue Musculoskeletal Diseases: From Molecular Basis to Therapy (Volume II))

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25 pages, 4466 KiB

Open AccessArticle

Biomanufacturing and Curcumin-Loading of Human Choroid Plexus Organoid-Derived Extracellular Vesicles from a Vertical-Wheel Bioreactor to Alleviate Neuro-Inflammation

by Justice Ene, Laureana Muok, Vanessa Gonzalez, Nicolas Sanchez, Aakash Nathani, Falak Syed, Zixiang Leonardo Liu, Mandip Singh, Tristan Driscoll and Yan Li

Biomedicines 2025, 13(5), 1069; https://doi.org/10.3390/biomedicines13051069 - 28 Apr 2025

Abstract

Background: Choroid plexus is a complex structure in the human brain that is responsible for the secretion of extracellular vesicles (EVs) in cerebrospinal fluid. Few studies to date have generated choroid plexus (ChP) organoids differentiated from human induced pluripotent stem cells (hiPSCs) and [...] Read more.

Background: Choroid plexus is a complex structure in the human brain that is responsible for the secretion of extracellular vesicles (EVs) in cerebrospinal fluid. Few studies to date have generated choroid plexus (ChP) organoids differentiated from human induced pluripotent stem cells (hiPSCs) and analyzed their secreted EVs. The scalable Vertical-Wheel bioreactors (VWBRs) provide low shear stress and a controlled environment. Methods: This study utilized VWBRs for the differentiation of hiPSCs into ChP organoids and generation of the secreted EVs compared to a static culture. Additionally, this study loaded curcumin into ChP organoid-derived EVs, performed EV lyophilization, and determined the ability of the re-hydrated EVs to alleviate neuro-inflammation. Results: The results demonstrated that the VWBR culture exhibited more aerobic metabolism and active glucose and glutamine consumption than the static control. Consequently, the ChP markers and Endosomal Sorting Complexes Required for Transport-dependent and -independent EV biogenesis genes were significantly upregulated (2–3-fold) in the VWBR, producing four-fold-higher EVs per mL media than the static control. The EVs retained similar size and zeta potential after lyophilization and re-hydration. The cells exposed to amyloid beta 42 oligomers and treated with the curcumin-loaded re-hydrated EVs showed high viability and the reduced inflammatory response determined by TNF-α and IL-6 expression. Conclusions: This study demonstrates a scalable bioreactor system to promote ChP organoid differentiation and generation of EV-based cell-free therapeutics to treat neural inflammation in various neurological disorders. Full article

(This article belongs to the Special Issue 3D Cell Culture Systems for Biomedical Research)

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16 pages, 5885 KiB

Open AccessArticle

Route of Application and Dose Evaluation of Dental Pulp Stem Cells for the Treatment of Sialadenitis Caused by Sjögren’s Syndrome: A Preclinical Study

by Zhihao Du, Lifang Feng, Yu Zhang, Xin Peng, Shan Zhang, Rui Zhao, Jia Lei, Xiaotong Li, Guangyan Yu and Chong Ding

Biomedicines 2025, 13(5), 1068; https://doi.org/10.3390/biomedicines13051068 - 28 Apr 2025

Abstract

Background: Sjögren’s syndrome (SS) is an autoimmune disorder characterized by sicca syndrome and/or systemic manifestations. In this study, non-obese diabetic (NOD) mice were used as an animal model for studying SS, to evaluate the optimal administration route and dose range of [...] Read more.

Background: Sjögren’s syndrome (SS) is an autoimmune disorder characterized by sicca syndrome and/or systemic manifestations. In this study, non-obese diabetic (NOD) mice were used as an animal model for studying SS, to evaluate the optimal administration route and dose range of dental pulp stem cells (DPSCs) in the treatment of sialadenitis caused by SS. Methods: Different doses of DPSCs were transplanted into the submandibular glands (SMGs) of 14-week-old NOD mice through two different methods: injection or retrograde perfusion through the catheter orifice into the SMG. At 21 weeks of age, the saliva flow rate (SFR), ectopic lymphocytes, and CD4⁺ T-cell infiltration were measured. Tumor necrosis factor-alpha (TNF-α) and interferon-gamma (IFN-γ) in the glandular tissues were also quantitatively detected. Results: Compared with untreated and PBS-injected controls, different-dose groups of the two administration methods showed an increased saliva flow rate of NOD mice to varying degrees, reduced infiltration of lymphocytes and CD4⁺ T cells in the SMG, and decreased IFN-γ/TNF-α levels. Finally, we compared these two administration routes and found that the perfusion of 2 × 10⁵ DPSCs presents good therapeutic effects. Conclusions: DPSC perfusion through the catheter orifice is a simple and effective treatment method, which is worthy of further investigation through clinical trials. Full article

(This article belongs to the Special Issue Pathogenesis, Diagnostics, and Therapeutics for Rheumatic Diseases)

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17 pages, 4142 KiB

Open AccessArticle

Acute Myeloid Leukemia Genome Characterization Study and Subtype Classification Employing Feature Selection and Bayesian Networks

by Zhenzhen Li, Jingwen Li, Sifan Li, Yangyang Wang and Jihan Wang

Biomedicines 2025, 13(5), 1067; https://doi.org/10.3390/biomedicines13051067 - 28 Apr 2025

Abstract

Background: The precise diagnosis and classification of acute myeloid leukemia (AML) has important implications for clinical management and medical research. Methods: We investigated the expression of protein-coding genes in blood samples from AML patients and controls using The Cancer Genome Atlas (TCGA) and [...] Read more.

Background: The precise diagnosis and classification of acute myeloid leukemia (AML) has important implications for clinical management and medical research. Methods: We investigated the expression of protein-coding genes in blood samples from AML patients and controls using The Cancer Genome Atlas (TCGA) and Genotype-Tissue Expression (GTEx) databases. Subsequently, we applied the feature selection method of the least absolute shrinkage and selection operator (LASSO) to select the optimal gene subset for classifying AML patients and controls as well as between a particular FAB subtype and other subtypes of AML. Results: Using LASSO method, we identified a subset of 101 genes that could effectively distinguish between AML patients and control individuals; these genes included 70 up-regulated and 31 down-regulated genes in AML. Functional annotation and pathway analysis indicated the involvement of these genes in RNA-related pathways, which was also consistent with the epigenetic changes observed in AML. Results from survival analysis revealed that several genes are correlated with the overall survival in AML patients. Additionally, LASSO-based gene subset analysis successfully revealed differences between certain AML subtypes, providing valuable insights into subtype-specific molecular mechanisms and differentiation therapy. Conclusions: This study demonstrated the application of machine learning in genomic data analysis for identifying gene subsets relevant to AML diagnosis and classification, which could aid in improving the understanding of the molecular landscape of AML. The identification of survival-related genes and subtype-specific markers may lead to the identification of novel targets for personalized medicine in the treatment of AML. Full article

(This article belongs to the Special Issue Advances in Genetic Research and Molecular Diagnostics for Hematological Diseases)

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16 pages, 297 KiB

Open AccessArticle

Challenges in the Treatment of Urinary Tract Infections: Antibiotic Resistance Profiles of Escherichia coli Strains Isolated from Young and Elderly Patients in a Southeastern Romanian Hospital

by Andreea-Elena Topa, Constantin Ionescu, Anca Pinzaru, Elena Mocanu, Ana Maria Iancu, Elena Dumea, Bogdan Florentin Nitu, Florin Gabriel Panculescu and Simona Claudia Cambrea

Biomedicines 2025, 13(5), 1066; https://doi.org/10.3390/biomedicines13051066 - 28 Apr 2025

Abstract

Background/Objectives: Urinary tract infections (UTIs) represent a significant public health challenge, with Escherichia coli being the primary causative pathogen. The rise in antimicrobial resistance (AMR), further intensified by shifts in antibiotic prescribing practices during and after the COVID-19 pandemic, poses substantial difficulties [...] Read more.

Background/Objectives: Urinary tract infections (UTIs) represent a significant public health challenge, with Escherichia coli being the primary causative pathogen. The rise in antimicrobial resistance (AMR), further intensified by shifts in antibiotic prescribing practices during and after the COVID-19 pandemic, poses substantial difficulties in treatment optimization and clinical management. Methods: This retrospective study analyzed 644 E. coli strains from urine samples collected in a southeastern Romanian hospital during two periods: pre-pandemic (2018–2019, N = 361) and post-pandemic (2023–2024, N = 283). Antimicrobial susceptibility was assessed using the VITEK automated system for key antibiotic classes. Results: A significant increase in fluoroquinolone resistance was observed, especially for ciprofloxacin (p = 0.02), alongside rising ceftriaxone resistance (p = 0.004), suggesting the spread of ESBL-producing strains. Resistance to trimethoprim/sulfamethoxazole, ampicillin, and amoxicillin/clavulanic acid remained high, limiting their empirical use. Carbapenem resistance was low (p > 0.1), while nitrofurantoin and fosfomycin retained high efficacy (p = 0.26 and p = 0.64). Conclusions: The post-pandemic period showed a concerning rise in resistance to fluoroquinolones and third-generation cephalosporins, highlighting the need for stricter antimicrobial stewardship. Carbapenems remain effective for severe infections, while nitrofurantoin and fosfomycin are reliable first-line options for uncomplicated UTIs. Continuous AMR surveillance is essential to optimize treatment and curb multidrug-resistant strains. Full article

(This article belongs to the Section Microbiology in Human Health and Disease)

19 pages, 4299 KiB

Open AccessArticle

Sinapic Acid Ameliorates Cadmium-Induced Hepatotoxicity: Modulation of Oxidative Stress, Inflammation, and Apoptosis

by Yomna A. Farahat, Norhan M. El-Sayed, Reem M. Hazem, Eman T. Mehanna and Asmaa Radwan

Biomedicines 2025, 13(5), 1065; https://doi.org/10.3390/biomedicines13051065 - 28 Apr 2025

Abstract

Background/Objectives: Cadmium (Cd) is a harmful metal commonly used in industry. Numerous clinical diseases, including osteomalacia, testicular damage, renal and hepatic failure, and pulmonary edema, are associated with Cd exposure. The current study evaluated the protective effect of Sinapic acid (SA) against [...] Read more.

Background/Objectives: Cadmium (Cd) is a harmful metal commonly used in industry. Numerous clinical diseases, including osteomalacia, testicular damage, renal and hepatic failure, and pulmonary edema, are associated with Cd exposure. The current study evaluated the protective effect of Sinapic acid (SA) against Cd-induced hepatotoxicity by investigating different mechanistic pathways interfering with Cd-related liver injury. Methods: Forty rats were randomly assigned to four groups as follows; group 1 served as negative control and received saline, group 2 received saline for 14 days and CdCl₂ (3.5 mg/kg IP) as a single dose on day 14, groups 3 and 4 were treated with SA (20, 40 mg/kg PO), respectively, for 14 days and injected with CdCl₂ (3.5 mg/kg IP) on day 14. Serum was collected to evaluate liver function. Liver samples were collected for histopathological examination and the assessment of markers related to oxidative stress, inflammation, and apoptosis. Results: Acute Cd administration elevated liver enzymes and induced pathological changes in liver specimens, with the concurrent release of inflammatory markers and reduced antioxidant capabilities. Pretreatment with SA improved liver function and Cd-induced histopathological changes and elevated the activities of antioxidant enzymes. SA ameliorated inflammation, as evidenced by decreased expression of NF-κB, TNF-α, TLR-4, and COX-2, iNOS, and IL-1β levels along with suppression of mTOR, JNK, ERK, BAX, and Bcl-2. Conclusions: The present data suggest that SA represents a promising protective agent against Cd-induced hepatic injury by attenuating oxidative stress, inflammation, and apoptosis. Full article

(This article belongs to the Section Drug Discovery, Development and Delivery)

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