Biomedicines

14 pages, 673 KiB

Open AccessArticle

Comparative Analysis of Rescue-In Vitro-Maturation (r-IVM) Outcomes in Women with Diminished Ovarian Reserve (DOR) Versus Normal Ovarian Reserve (NOR)

by Mohd Faizal Ahmad, Nurul Yaqin Mohd Nor, Mohammad Mahmoud Mohammad Ramadneh, Nurul Izyani Roseli, Marjanu Hikmah Elias, Norazilah Mat Jin, Muhammad Azrai Abu, Saiful Effendi Syafruddin, Ani Amelia Zainuddin, Shah Shamsul Azhar, Nao Suzuki and Abdul Kadir Abdul Karim

Biomedicines 2025, 13(5), 1084; https://doi.org/10.3390/biomedicines13051084 (registering DOI) - 29 Apr 2025

Abstract

Background/Objectives: Diminished ovarian reserve (DOR) poses significant challenges in the reproductive field, resulting in fewer mature and more low-quality eggs. Methods: We studied r-IVM in addition to standard in vitro fertilization (IVF) and compared the embryological outcomes between both DOR and NOR women. [...] Read more.

Background/Objectives: Diminished ovarian reserve (DOR) poses significant challenges in the reproductive field, resulting in fewer mature and more low-quality eggs. Methods: We studied r-IVM in addition to standard in vitro fertilization (IVF) and compared the embryological outcomes between both DOR and NOR women. Results: We recruited 90 women (45 NOR; 45 DOR) with a younger age seen in NOR (35.2 vs. 36.5 years old) women. Otherwise, DOR women had lower levels of AMH and AFC, thus fewer retrieved follicles and collected oocytes. Most of the group presented with primary subfertility, with 55.6% in the NOR group diagnosed with polycystic ovary syndrome (PCOS), while 37.8% in the DOR group presented with aging and cancer survivorship issues. Most women in the NOR group used hCG as a trigger (82.2%), while 17.8% of the DOR group opted for a decapeptide. A total of 719 oocytes were retrieved, with 72.3% of eggs being mature in the NOR group compared to 64.9% in the DOR group. Following r-IVM, 47.69% of NOR eggs were matured compared to 60% in DOR eggs. The fertilization rates (FRs) following r-IVM were higher in the DOR group (66.7% vs. 37.8%). Overall, higher numbers and quality of D3 embryos were seen in the DOR group. Our analysis revealed that the trigger type, hCG, was the only significant factor linked to successful oocyte maturation rates. Conclusions: Our study suggests that r-IVM may enhance outcomes for women with DOR, including better egg maturity, FR, and embryo quality than NOR women. Full article

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

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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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

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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

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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

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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

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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

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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

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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

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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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16 pages, 3287 KiB

Open AccessArticle

Evaluating Magnetic Stimulation as an Innovative Approach for Treating Dry Eye Disease: An Initial Safety and Efficacy Study

by Hadas Ben-Eli, Shimon Perelman, Denise Wajnsztajn and Abraham Solomon

Biomedicines 2025, 13(5), 1064; https://doi.org/10.3390/biomedicines13051064 - 28 Apr 2025

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Abstract

Objective: The aim of this study was to assess the safety and preliminary efficacy of repetitive magnetic stimulation (RMS) as a treatment intervention for dry eye disease (DED), focusing on symptom reduction. Methodology: This investigation involved 22 adult participants (85% females, aged between [...] Read more.

Objective: The aim of this study was to assess the safety and preliminary efficacy of repetitive magnetic stimulation (RMS) as a treatment intervention for dry eye disease (DED), focusing on symptom reduction. Methodology: This investigation involved 22 adult participants (85% females, aged between 22 and 79 years) diagnosed with moderate-to-severe DED. These individuals were subjected to RMS treatment targeting one or both eyes using the VIVEYE-Ocular Magnetic Neurostimulation System version 1.0 (Epitech-Mag LTD; National Institute of Health (NIH) clinical trials registry #NCT03012698). A placebo-controlled group was also included for comparative analysis, with all subjects being monitored over a three-month period. The evaluation of safety encompassed monitoring changes in best corrected visual acuity, ocular pathology, and the reporting of adverse events. Participant tolerance was gauged through questionnaires, measurements of intraocular pressure (IOP), Schirmer’s test, and vital signs. The efficacy of the treatment was assessed by comparing pre- and post-treatment scores for fluorescein staining (according to National Eye Institute (NEI) grading) and patient-reported outcomes. Results: No statistically significant changes were found in visual acuity, IOP, or Schirmer’s test results between the RMS-treated and control groups (p < 0.05), indicating that RMS does not negatively impact these ocular functions. However, RMS treatment was associated with improved tear film stability (p = 0.19 vs. p = 0.04) and corneal health (p = 0.52 vs. p = 0.004), with no improvements in the control group. Initial symptom improvement was observed in both RMS-treated and placebo groups (p = 0.007 vs. p = 0.008), suggesting a potential therapeutic benefit of RMS for ocular surface conditions beyond a placebo effect. Conclusions: This study presents RMS as a promising therapeutic approach for DED, highlighting its potential to promote corneal epithelial repair, enhance tear film stability, and improve patient-reported symptoms without negatively impacting IOP, visual acuity, or tear production. This confirms the safety and suggests the efficacy of RMS therapy for dry eye conditions. Full article

(This article belongs to the Special Issue Recent Research on Dry Eye)

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10 pages, 3180 KiB

Open AccessArticle

Clinically Uncertain Liver Masses: A Guide to Distinguishing Poorly Differentiated Primary Liver Cancer

by Greta Sökeland, Michael P. Brönnimann, Erik Vassella, Guido Stirnimann, Matteo Montani and Juliane Friemel

Biomedicines 2025, 13(5), 1063; https://doi.org/10.3390/biomedicines13051063 - 28 Apr 2025

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Abstract

Objectives: The distinction of clinically uncertain, poorly differentiated liver masses into primary liver cancer (PLC) of cholangiocytic origin (intrahepatic cholangiocarcinoma; CCA) or hepatocellular origin (hepatocellular carcinoma; HCC) vs. metastasis is highly relevant to guiding patient treatment. Protocols differ in terms of resection, [...] Read more.

Objectives: The distinction of clinically uncertain, poorly differentiated liver masses into primary liver cancer (PLC) of cholangiocytic origin (intrahepatic cholangiocarcinoma; CCA) or hepatocellular origin (hepatocellular carcinoma; HCC) vs. metastasis is highly relevant to guiding patient treatment. Protocols differ in terms of resection, local ablation, liver transplantation, or systemic therapies with immune checkpoint inhibitors (ICIs) and tyrosine kinase inhibitors (TKIs). Methods: This retrospective case series exemplifies a multidisciplinary, practical guide to clinically uncertain liver masses using imaging, histomorphology, immune phenotyping, and mutational testing of telomerase promoter (TERT) combined with a literature review. Results: In 2/3 patients with uncertain liver masses and inconclusive immunohistochemistry profiles, TERT testing supported the diagnosis of poorly differentiated hepatocellular carcinoma. The third case with a history of sclerosing cholangitis and vague adenoid morphology yielded mutations in ARID1a and TP53 and was identified as primary liver cancer, consistent with poorly differentiated intrahepatic cholangiocarcinoma or mixed hepatocellular cholangiocarcinoma (cHCC/CCA). Conclusions: Finding HCC-typical TERT promoter mutations is a useful diagnostic tool in poorly differentiated primary liver cancer. Full article

(This article belongs to the Special Issue Advanced Cancer Diagnosis and Treatment: Second Edition)

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22 pages, 7330 KiB

Open AccessArticle

Relevance of Cellular Homeostasis-Related Gene Expression Signatures in Distinct Molecular Subtypes of Breast Cancer

by Sharda P. Singh, Chathurika S. Dhanasekara, Michael W. Melkus, Chhanda Bose, Sonia Y. Khan, Flavia Sardela de Miranda, Maria F. Mahecha, Prrishti J. Gukhool, Sahil S. Tonk, Se-Ran Jun, Sahra Uygun and Rakhshanda Layeequr Rahman

Biomedicines 2025, 13(5), 1058; https://doi.org/10.3390/biomedicines13051058 - 28 Apr 2025

Viewed by 134

Abstract

Background: Breast cancer is a complex and heterogeneous disease characterized by distinct molecular subtypes with varying prognoses and treatment responses. Multiple factors influence breast cancer outcomes including tumor biology, patient characteristics, and treatment modalities. Demographic factors such as age, race/ethnicity, menopausal status, and [...] Read more.

Background: Breast cancer is a complex and heterogeneous disease characterized by distinct molecular subtypes with varying prognoses and treatment responses. Multiple factors influence breast cancer outcomes including tumor biology, patient characteristics, and treatment modalities. Demographic factors such as age, race/ethnicity, menopausal status, and body mass index have been correlated with variations in incidence, mortality, and survival rates. Over the past decade, comprehensive genomic profiling has been widely used to identify molecular biomarkers and signatures to develop novel therapeutic strategies for patients. For instance, the FLEX registry (NCT03053193) enrolled stage I–III breast cancer patients across 90 institutions in the United States and stratified risk groups based on a 70-gene signature (MammaPrint^®-MP) and molecular subtype based on an 80-gene signature (BluePrint^®-BP). This study aimed to identify the gene expression patterns and biomarkers associated with breast cancer risk and progression by integrating transcriptomic and clinical data. Methods: Targeted 111 unique gene expression and clinical data points from 978 breast cancer samples, representing each BP subtype (26% Luminal A, 26% Luminal B, 25% Basal, 23% HER2), obtained from Agendia Inc. These genes were selected based on their involvement in the mercapturic acid pathway, white and brown adipose tissue markers, inflammation markers, tumor-associated genes, apoptosis, autophagy, and ER stress markers. All statistical analyses, including principal component analysis (PCA), were performed using R version [4.4.0]. Prognostic values and genetic alterations were investigated using various web-based programs as described in the Methods section. Results: PCA of gene expression data revealed distinct clustering patterns associated with risk categories and molecular subtypes, particularly with principal component 4 (PC4). Genes related to oxidative stress, autophagy, apoptosis, and histone modification showed altered expression across risk categories and molecular subtypes. Key differentially expressed genes included SOD2, KLK5, KLK7, IL8, GSTM1/2, GLI1, CBS, and IGF1. Pathway analysis highlighted the enrichment of processes related to autophagy, cellular stress response, apoptosis, glutathione metabolism, deacetylation, and oxidative stress in high-risk and basal-like tumors compared with Ultralow and Luminal A tumors, respectively. Conclusions: This study identified gene expression signatures associated with breast cancer risk and molecular subtypes. These findings provide insights into the biological processes that may drive breast cancer progression and could inform the development of prognostic biomarkers and personalized therapeutic strategies. Full article

(This article belongs to the Special Issue Triple Negative Breast Cancer: From Mechanisms to Therapeutic Approaches)

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14 pages, 604 KiB

Open AccessSystematic Review

Clinical Molecular Immunohistochemistry Mismatch Repair Mutations in Lynch Syndrome in Patients Under 50 Years: A Systematic Review

by Bogdan Adrian Manta, Adrian Cosmin Ilie, Felicia Marc, Daciana Nistor, Patricia Octavia Mazilu and Claudia Borza

Biomedicines 2025, 13(5), 1062; https://doi.org/10.3390/biomedicines13051062 - 27 Apr 2025

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Abstract

Background and Objectives: Lynch syndrome (LS), an autosomal dominant condition arising from germline mutations in mismatch repair (MMR) genes, is a major cause of hereditary early-onset colorectal cancer (CRC). Although patients diagnosed before age 50 represent a critical subgroup where Lynch syndrome [...] Read more.

Background and Objectives: Lynch syndrome (LS), an autosomal dominant condition arising from germline mutations in mismatch repair (MMR) genes, is a major cause of hereditary early-onset colorectal cancer (CRC). Although patients diagnosed before age 50 represent a critical subgroup where Lynch syndrome might be more prevalent, data on the precise frequency, clinical outcomes, and molecular correlates remain heterogeneous across studies. This systematic review was conducted to (1) estimate the prevalence of MMR deficiency (dMMR) and confirmed LS in patients diagnosed with CRC before the age of 50, and (2) examine immunohistochemistry (IHC) mismatch repair testing patterns and associated molecular findings (BRAF mutations, MLH1 promoter hypermethylation, somatic MMR gene alterations). Methods: Following a predefined search strategy in PubMed, Scopus, and Web of Science, five relevant studies were identified (n = 5). Each study comprised patients younger than 50 who underwent IHC-based tumor screening. Data extraction covered demographic details, number of patients tested, proportion with abnormal IHC, frequency of somatic or germline MMR gene mutations, and method of classification into sporadic dMMR vs. LS. Quality assessment was performed using recommended scales for observational studies. Results: Among 5 studies totaling 960 early-onset CRC patients, the frequency of dMMR CRC ranged from 8.4% to 19.1%. The confirmed prevalence of LS among all young-onset CRC was between 5.0% and 5.9% in three studies but reached 8.9% in another and 5.1% in yet another. Across all studies, the presence of right-sided tumors and histopathological features such as tumor-infiltrating lymphocytes were more common in dMMR cancers. Incorporation of MLH1-promoter hypermethylation and/or BRAF V600E mutation testing aided discrimination of sporadic dMMR CRC from germline LS cases. Conclusions: The prevalence of LS in CRC patients younger than 50 is clinically significant, at approximately 5–9%. Routine IHC-based MMR screening is both feasible and effective for detecting LS in early-onset CRC. Further research is needed to standardize universal testing protocols, delineate the role of additional molecular assays, and ensure comprehensive genetic counseling for at-risk individuals. Full article

(This article belongs to the Section Immunology and Immunotherapy)

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12 pages, 1069 KiB

Open AccessArticle

Perimesencephalic Subarachnoid Hemorrhage Is Not Always a Benign Condition: Hemorrhage Volume as a Predictor for Complications and Clinical Outcome

by Emily Hoffmann, Công Duy Bùi, Alexandra Valls Chavarria, Michael Müther, Markus Holling, Manfred Musigmann, Max Masthoff, Mostafa Ergawy, Tobias D. Faizy, Christian Paul Stracke, Hermann Krähling and Burak Han Akkurt

Biomedicines 2025, 13(5), 1061; https://doi.org/10.3390/biomedicines13051061 - 27 Apr 2025

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Abstract

Objective: The benign nature of perimesencephalic subarachnoid hemorrhage (pmSAH) can be challenged by the occurrence of complications. Given the limited prognostic value of established clinical parameters for the development of complications in patients with pmSAH, this study evaluates the potential of volumetric hemorrhage [...] Read more.

Objective: The benign nature of perimesencephalic subarachnoid hemorrhage (pmSAH) can be challenged by the occurrence of complications. Given the limited prognostic value of established clinical parameters for the development of complications in patients with pmSAH, this study evaluates the potential of volumetric hemorrhage quantification for risk assessment and the evaluation of the clinical outcome. Material and Methods: In this retrospective single-center study, we analyzed all consecutive patients diagnosed with pmSAH between 2010 and 2023 at a tertiary care academic medical center in Germany. The volumetric quantification of the hemorrhage in cm³ was performed using non-contrast CT imaging. The occurrence of clinical complications, including hydrocephalus, vasospasm, and delayed cerebral ischemia (DCI), were assessed. Clinical outcomes were determined by the Glasgow Outcome Scale (GOS) at discharge. Multivariable logistic regression models were used to assess the correlation between quantified hemorrhage volumes and the occurrence of complications and clinical outcomes (GOS) controlled for other variables such as age, sex, cardiovascular risk factors, clinical symptoms, and the modified Fisher scale. Results: A total of 82 patients (58.5% male, 54.8 ± 12.1 years) were enrolled. The median World Federation of Neurosurgical Societies (WFNS) score for all patients at admission was 1.0 (IQR 1.0–1.0). During the clinical course, hydrocephalus occurred in 29%, vasospasm in 14.6%, and DCI in 8.5% of all patients. Hemorrhage volume quantification was found to be the strongest independent predictor for hydrocephalus (OR 1.28; 95% CI 1.02–1.61; p = 0.032) and vasospasm (OR 1.25; 95% CI 1.07–1.46; p = 0.007) and showed a high predictive accuracy in ROC analyses (AUC = 0.77 and 0.76, respectively). Conversely, neither clinical parameters nor the modified Fisher scale were associated with these complications. A higher hemorrhage volume was also significantly correlated with a worse functional outcome (GOS; β = –0.07, CI: −0.12–−0.02, p = 0.021). Conclusions: In patients with pmSAH, the volumetric quantification of hemorrhage may be an adequate prognostic parameter regarding the occurrence of hydrocephalus and vasospasm. In addition, the quantitative assessment of hemorrhage volumes was strongly associated with clinical outcomes in these patients. Despite the generally benign nature of pmSAH, this imaging biomarker could improve individualized clinical management strategies and inform about the risk for the occurrence of complications. Full article

(This article belongs to the Special Issue Emerging Biomarkers and Therapeutic Approaches in Neurovascular Diseases)

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15 pages, 7732 KiB

Open AccessArticle

Construction of Diagnostic Model for Regulatory T Cell-Related Genes in Sepsis Based on Machine Learning

by Xuesong Wang, Zhe Guo, Xinrui Wang and Zhong Wang

Biomedicines 2025, 13(5), 1060; https://doi.org/10.3390/biomedicines13051060 - 27 Apr 2025

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Abstract

Background: Sepsis is a complex syndrome caused by a severe infection that occurs with a severe inflammatory response. Regulatory T cells (Tregs) have immunosuppressive effects and play a crucial role in modulating the immune response. There-fore, the number of Tregs is significantly increased [...] Read more.

Background: Sepsis is a complex syndrome caused by a severe infection that occurs with a severe inflammatory response. Regulatory T cells (Tregs) have immunosuppressive effects and play a crucial role in modulating the immune response. There-fore, the number of Tregs is significantly increased in sepsis patients. Methods and Results: This paper aims to identify Tregs associated with the diagnosis of sepsis. For this purpose, transcriptional data from the GEO database for sepsis and its controls were downloaded and subjected to differential expression analysis. Immuno-infiltration analysis of the obtained DEGs revealed that Tregs were significantly different in sepsis and its controls. To further explore the cellular landscape and interactions in sepsis, single-cell RNA sequencing (scRNA-seq) data were analyzed. We identified key cell types and their interactions, including Tregs, using cell–cell communication analysis tools such as CellChat. This analysis provided in-sights into the dynamic changes in immune cell populations and their communication networks in sepsis. Thus, we utilized multiple machine learning algorithms to screen and extract Treg-related genes associated with sepsis diagnosis. We then performed both in-ternal and external validation tests. The final diagnostic model was constructed with high diagnostic accuracy (accuracy of 0.9615). Furthermore, we verified the diagnostic gene via a qPCR experiment. Conclusions: This paper elucidates the potential diagnostic targets associated with Tregs in sepsis progression and provides comprehensive understanding of the immune cell interactions in sepsis through scRNA-seq analysis. Full article

(This article belongs to the Collection Feature Papers in Immunology and Immunotherapy)

24 pages, 4202 KiB

Open AccessArticle

Resveratrol-Loaded Solid Lipid Nanoparticles Reinforced Hyaluronic Hydrogel: Multitarget Strategy for the Treatment of Diabetes-Related Periodontitis

by Raffaele Conte, Anna Valentino, Fabrizia Sepe, Francesco Gianfreda, Roberta Condò, Loredana Cerroni, Anna Calarco and Gianfranco Peluso

Biomedicines 2025, 13(5), 1059; https://doi.org/10.3390/biomedicines13051059 - 27 Apr 2025

Viewed by 154

Abstract

Background/Objectives: Periodontitis and diabetes mellitus share a well-established bidirectional relationship, where hyperglycemia exacerbates periodontal inflammation, and periodontal disease further impairs glycemic control. Within the diabetic periodontal microenvironment, an imbalance between pro-inflammatory (M1) and anti-inflammatory (M2) macrophages promotes chronic inflammation, oxidative stress, delayed healing, [...] Read more.

Background/Objectives: Periodontitis and diabetes mellitus share a well-established bidirectional relationship, where hyperglycemia exacerbates periodontal inflammation, and periodontal disease further impairs glycemic control. Within the diabetic periodontal microenvironment, an imbalance between pro-inflammatory (M1) and anti-inflammatory (M2) macrophages promotes chronic inflammation, oxidative stress, delayed healing, and alveolar bone resorption. Resveratrol (RSV), a polyphenol with antioxidant, anti-inflammatory, and pro-osteogenic properties, holds potential to restore macrophage balance. However, its clinical application is limited by poor bioavailability and instability. This study aimed to develop and evaluate a novel RSV delivery system to overcome these limitations and promote periodontal tissue regeneration under diabetic conditions. Methods: A drug delivery system comprising RSV-loaded solid lipid nanoparticles embedded within a cross-linked hyaluronic acid hydrogel (RSV@CLgel) was formulated. The system was tested under hyperglycemic and inflammatory conditions for its effects on macrophage polarization, cytokine expression, oxidative stress, mitochondrial function, and osteoblast differentiation. Results: RSV@CLgel effectively suppressed pro-inflammatory cytokines (TNF-α, IL-1β, IL-6) while upregulating anti-inflammatory markers (IL-10, TGF-β). It significantly reduced oxidative stress by decreasing ROS and lipid peroxidation levels and improved mitochondrial function and antioxidant enzyme activity. Furthermore, RSV@CLgel enhanced osteoblast differentiation, as evidenced by increased ALP activity, calcium nodule formation, and upregulation of osteogenic genes (COL-I, RUNX2, OCN, OPN). It also inhibited RANKL-induced osteoclastogenesis, contributing to alveolar bone preservation. Conclusions: The RSV@CLgel delivery system presents a promising multifunctional strategy for the management of diabetic periodontitis. By modulating immune responses, reducing oxidative stress, and promoting periodontal tissue regeneration, RSV@CLgel addresses key pathological aspects of diabetes-associated periodontal disease. Full article

(This article belongs to the Special Issue Periodontal Disease and Periodontal Tissue Regeneration)

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15 pages, 539 KiB

Open AccessReview

G-Quadruplexes in Tumor Immune Regulation: Molecular Mechanisms and Therapeutic Prospects in Gastrointestinal Cancers

by Yunxia Zhou, Difei Xu, Ying Zhang and Huaixiang Zhou

Biomedicines 2025, 13(5), 1057; https://doi.org/10.3390/biomedicines13051057 - 27 Apr 2025

Viewed by 154

Abstract

G-quadruplex (G4) is a noncanonical nucleic acid secondary structure self-assembled by guanine-rich sequences. Recent studies have not only revealed the key role of G4 in gene regulation, DNA replication, and telomere maintenance but also showed that it plays a core role in regulating [...] Read more.

G-quadruplex (G4) is a noncanonical nucleic acid secondary structure self-assembled by guanine-rich sequences. Recent studies have not only revealed the key role of G4 in gene regulation, DNA replication, and telomere maintenance but also showed that it plays a core role in regulating the tumor immune microenvironment. G4 participates in tumor immune escape and the inhibition of immune response by regulating immune checkpoint molecules, cytokine expression, immune cell function, and their interaction network, thus significantly affecting the effect of tumor immunotherapy. This article systematically reviews the molecular mechanism of G4 in tumor immune regulation, especially gastrointestinal tumors, and explores the potential and application prospects of G4-targeted drug strategies in improving anti-tumor immunotherapy. 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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17 pages, 3030 KiB

Open AccessArticle

Pharmacological Blocking of Adiponectin Receptors Induces Alzheimer’s Disease-like Neuropathology and Impairs Hippocampal Function

by Hui-Hui Guo, Hai-Ning Ou, Jia-Sui Yu, Suk-Yu Yau and Hector Wing-Hong Tsang

Biomedicines 2025, 13(5), 1056; https://doi.org/10.3390/biomedicines13051056 - 27 Apr 2025

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Abstract

Background/Objectives: Previous studies have shown that adiponectin deficiency or blocking adiponectin receptors (AdipoRs) in the brain can lead to an Alzheimer’s disease (AD)-like neuropathology. While AdipoRs are abundantly expressed in peripheral tissues, the effects of blocking these receptors in the peripheral tissues [...] Read more.

Background/Objectives: Previous studies have shown that adiponectin deficiency or blocking adiponectin receptors (AdipoRs) in the brain can lead to an Alzheimer’s disease (AD)-like neuropathology. While AdipoRs are abundantly expressed in peripheral tissues, the effects of blocking these receptors in the peripheral tissues on the brain are unclear. This study investigates the impacts of blocking AdipoRs with a peripheral administration of ADP400, an antagonist peptide that targets AdipoRs on cognitive performance, hippocampal adult neurogenesis, and AD-like neuropathology in mice. Methods: Adult mice were intraperitoneally administered with ADP400 peptide that blocks peripheral AdipoRs continuously for 21 days, followed by a battery of behavioral test for mood and memory performance. Results: ADP400-treated mice exhibited impaired memory performance and increased anxiety-like behaviors. Molecular analyses revealed heightened hyperphosphorylation of tau and increased β-amyloid levels, alongside decreased expression of AdipoRs and PP2A in the hippocampus, suggesting a critical role of AdipoRs in AD-like neuropathology. Furthermore, ADP400 treatment significantly reduced hippocampal adult neurogenesis, as indicated by decreased BrdU, Ki67, and DCX staining. Inhibiting peripheral adiponectin receptors could lead to tau hyperphosphorylation and accumulated β-amyloid levels. Conclusions: These findings highlight the critical role of peripheral manipulation of adiponectin receptors in modulating cognitive function and adult neurogenesis, offering insights into potential therapeutic strategies for AD and related cognitive disorders. Full article

(This article belongs to the Special Issue Recent Advances in Adipokines—2nd Edition)

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14 pages, 2450 KiB

Open AccessArticle

Bleomycin Electrosclerotherapy (BEST) for Slow-Flow Malformations of the Upper Aerodigestive Tract

by Veronika Vielsmeier, Vanessa F. Schmidt, Florian Obereisenbuchner, Natascha Platz Batista da Silva, Walter A. Wohlgemuth, Daniel Puhr-Westerheide, Max Seidensticker, Jens Ricke, Thomas Kühnel, Christopher Bohr, Moritz Wildgruber and Caroline T. Seebauer

Biomedicines 2025, 13(5), 1055; https://doi.org/10.3390/biomedicines13051055 - 27 Apr 2025

Viewed by 103

Abstract

Background/Objectives: Bleomycin electrosclerotherapy (BEST), which combines intralesional bleomycin administration with electroporation, enhances drug uptake and has shown efficacy in treating vascular malformations resistant to conventional therapies. While BEST is increasingly used in various anatomical sites, its application in the upper aerodigestive tract remains [...] Read more.

Background/Objectives: Bleomycin electrosclerotherapy (BEST), which combines intralesional bleomycin administration with electroporation, enhances drug uptake and has shown efficacy in treating vascular malformations resistant to conventional therapies. While BEST is increasingly used in various anatomical sites, its application in the upper aerodigestive tract remains underexplored. This study evaluates the safety and effectiveness of BEST in managing slow-flow vascular malformations of the oral cavity, tongue, larynx, and hypopharynx. Methods: In this retrospective, multicenter study, 20 patients with symptomatic slow-flow vascular malformations of the upper aerodigestive tract were treated with BEST. Clinical and radiological assessments were used to evaluate the treatment response, categorized as “significantly reduced”, “reduced”, “stable disease”, or “lesion growth”. Postprocedural complications and functional outcomes were systematically recorded. Results: A total of 29 BEST sessions were performed. Lesions of the tongue (n = 8) and combined oral cavity and tongue (n = 6) showed the highest response rates, with significant symptom reduction in five out of eight and five out of six patients, respectively. Among isolated oral cavity lesions (n = 4), one out of four demonstrated a significant reduction. In contrast, laryngeal and hypopharyngeal lesions (n = 2) had limited response, with one case showing partial reduction and the other remaining stable. Severe complications, including bleeding and dyspnea requiring tracheostomy, limited further treatment in these locations. No systemic adverse events, such as pulmonary toxicity, were observed. Conclusions: BEST is effective for treating vascular malformations of the upper aerodigestive tract, particularly in the tongue and oral cavity, but presents significant risks in laryngeal and hypopharyngeal lesions. A multidisciplinary approach is required to optimize treatment protocols for these challenging locations. Full article

(This article belongs to the Special Issue New Perspectives in the Treatment of Head and Neck Vascular Malformations)

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Biomedicines, Volume 13, Issue 5 (May 2025) – 76 articles

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