Abstracts of the 3rd International Electronic Conference on Biomedicines ^†

Abstract

The 3rd International Electronic Conference on Biomedicines (ECB 2025) was an online event held from 12 to 15 May 2025, by the journal Biomedicines MDPI. The goal of this conference was to enable scientists to present their latest research in the field of biomedicines, especially on the discovery and characterization of new therapeutic targets in the era of precise medicine, and to engage in discussions with the scientific community worldwide. Recent advances in biomedical research have provided an advanced insight into the biological background of human disease, setting the groundwork for the development of new therapeutic strategies. The agenda of the conference included, but was not limited to, the following topics: insulin resistance or hyperinsulinemia; immunotherapy and adverse effects; the tumor microenvironment; the immune system, tumor immunology, and autoimmune disease; biomedicine in cardiovascular diseases; diabetes, obesity, and metabolic diseases; gene therapy and gene editing; rare diseases; and molecular mechanisms of neurodegeneration.

1. Insulin Resistance or Hyperinsulinemia

1.1. Hypoglycemic Activity of New S-Derivatives of 6-(5-Mercapto-4-methyl-4H-1,2,4-triazol-3-yl)pyrimidine-2,4(1H,3H)-dione

Yuriy Karpenko

• Department of toxicological and inorganic chemistry, 1 Faculty of Pharmacy, Zaporizhzhia State Medical and Pharmaceutical University, Zaporizhzhia 69014, Ukraine

A fundamental aspect of the pathophysiology of type 2 diabetes is the progressive decline in the body’s response to prandial insulin, particularly in the early stages of the disease. The early-phase insulin response is significantly impaired, playing a key role in accelerating endogenous glucose production. In type 2 diabetes, endogenous glucose production persists despite prandial glucose intake. Combined with a persistent relative deficiency in insulin secretion, this leads to a predisposition to postprandial hyperglycemia.

The structural motif 1,2,4-Triazole is frequently explored in the development of new compounds with potential therapeutic applications, particularly for diabetes mellitus and other metabolic disorders. These compounds can modulate the enzymes involved in glucose metabolism, including AMP-activated protein kinase (AMPK).

The potential hypoglycemic activity of a new compound was evaluated by measuring changes in blood glucose levels in animals after a single administration. Male Wistar rats (180–200 g) were used as test subjects and were housed under standard vivarium conditions. The animals were divided into two groups of six (n = 6). The control group received a third-generation sulfonylurea derivative, glimepiride (4 mg/kg), a structurally related hypoglycemic drug. The test group received the experimental compound orally via gavage, either as an aqueous solution or as a finely dispersed aqueous emulsion stabilized with Tween-80. Blood samples were collected from the tail vein before administration and at 2, 4, 6, and 8 h post-administration.

Conclusions: The study results demonstrated that 2-((5-(2,6-dioxo-1,2,3,6-tetrahydropyrimidin-4-yl)-4-ethyl-4H-1,2,4-triazol-3-yl)thio)acetamide exhibits 1.5 times greater hypoglycemic activity than glimepiride. Molecular docking analysis confirmed the compound’s high affinity for the α-glucosidase enzyme (PDB ID: 2ZE0).

1.2. The Role of Insulin Resistance in Congestive Heart Failure, Impaired Glucose Tolerance, and Hypertension

Ishant Diwakar Dahake

• Department of Pharmaceutical Sciences, Rashtrasant Tukadoji Maharaj Nagpur University, Nagpur, 440033, India

Aim: The aim of this study was to explore the role of insulin resistance and compensatory hyperinsulinemia in glucose intolerance, hypertension, and CVD across conditions such as congestive heart failure (CHF) and non-insulin-dependent diabetes mellitus (NIDDM).

Background: Insulin resistance is a characteristic feature of congestive heart failure (CHF), non-insulin-dependent diabetes mellitus (NIDDM), and essential hypertension. Hyperinsulinemia, increased plasma norepinephrine, and disturbed glucose metabolism contribute to these conditions. Although compensatory hyperinsulinemia maintains glucose homeostasis, it may enhance cardiovascular risks by promoting endothelial dysfunction and dyslipidemia.

Methodology: The analysis synthesizes findings from the following:

• A case–control study of congestive heart failure patients and controls, using oral glucose tolerance tests and euglycemic clamps.
• Longitudinal studies of insulin-mediated glucose uptake and β-cell function in NIDDM and hypertension.
• Population-based studies of fasting insulin, norepinephrine, and free fatty acids (FFAs).

Results and Discussion: CHF patients presented with increased levels of norepinephrine, insulin, and FFA. These resulted in impaired glucose clearance and hepatic glucose regulation via insulin. NIDDM required hyperinsulinemia to counteract insulin resistance and maintain glucose tolerance, ultimately failing as the function of the β-cells declined. Hypertension linked insulin resistance with dyslipidemia and CVD, characterized by high triglycerides and low HDL cholesterol. Insulin resistance in CHF and hypertension further exacerbates metabolic derangement and increases the CVD risk.

Conclusions: The mechanisms of insulin resistance and compensatory hyperinsulinemia are central to the pathogenesis of glucose intolerance, hypertension, and CVD in CHF, NIDDM, and related disorders. Addressing these disturbances may help mitigate disease progression and improve cardiovascular outcomes.

2. Immunotherapy and Adverse Effects

2.1. A New, Edible, Plant-Derived Toxin as a Potential Candidate for Immunotherapy: An Evaluation of the Cytotoxic Profile and Its Effects on Intestinal Cells

Francesco Biscotti ^1,*, Federica Falà ^1,*, Massimo Bortolotti ¹, Sara Ragucci ², Nicola Landi ^2,3, Antimo Di Maro ², Andrea Bolognesi ¹, Letizia Polito ¹

^1. 

Department of Medical and Surgical Sciences (DIMEC), Alma Mater Studiorum University of Bologna, 40126 Bologna, Italy

^2. 

Department of Environmental, Biological and Pharmaceutical Sciences and Technologies (DiSTABiF), University of Campania ‘Luigi Vanvitelli’, Via Vivaldi 43, 81100 Caserta, Italy

^3. 

Institute of Crystallography, National Research Council of Italy, Via Vivaldi 43, 81100 Caserta, Italy

Introduction: Ribosome-inactivating proteins (RIPs) are toxic plant enzymes. Several RIPs have been conjugated to specific carriers to obtain immunotoxins, which gave promising results in preclinical and clinical studies. However, some adverse effects were reported, mainly immunogenicity. Recently, a new RIP from the edible plant Salsola soda, named sodin-5, has been identified and characterized. The aim of this work was to evaluate the sodin-5 cytotoxic profile and its effects on intestinal cell lines, comparing it with saporin, one of the best known and most studied RIPs for the construction of immunotoxins.

Methods: Different endpoints were considered to evaluate the RIPs’ cytotoxicity in colon adenocarcinoma Caco-2 and HT29 cell lines: cell viability, assessed through tetrazolium salt reduction, and cell death, assessed through cytofluorimetric analyses of AnnexinV/PI positivity. The effect of the RIPs on the membrane integrity was monitored by Trans-Epithelial Electrical Resistance (TEER) measurements on a Caco-2 monoculture and/or a Caco-2/HT29 co-culture. The RIPs’ immunological cross-reactivity was evaluated through ELISA.

Results: Our results showed that sodin-5 had high cytotoxicity, very similar to that of saporin. Both RIPs reduced cell viability after 72 h, with EC50s in the nM range. Apoptosis was the main cell death pathway triggered by the RIPs, and no necrosis involvement was observed. The lack of necrosis is very important as apoptosis does not cause inflammation. Sodin-5 and saporin significantly reduced the TEER values, both in the Caco-2 cell monoculture and the Caco-2/HT29 co-culture, in comparison to the untreated controls. Finally, no immunological cross-reactivity was observed with a saporin anti-serum.

Conclusions: Since sodin-5 and saporin have similar cytotoxic activities, sodin-5 could be proposed for further study as a potential toxic payload in immunotoxin construction to overcome some adverse effects of saporin and other RIPs. Theoretically, RIPs from edible sources should be less immunogenic and better tolerated by humans; therefore, sodin-5 could represent a better candidate for immunotoxin-based experimental therapy.

2.2. Combined In Vivo and In Cellulo Approaches to Study the Role of Endothelial Cells and the NLRP3 Inflammasome in Cardiac Dysfunction Associated with Immune-Mediated Myocarditis

Simon LLEDO ¹, Samantha Conte ¹, Thi-Thom Tran ^1,2, Juliette Vahdat ³, Joseph Ciccolini ⁴, Emmanuel Fenouillet ¹, Denis Puthier ⁵, Franck Thuny ^1,6,7, Jennifer Cautela ^1,6,7, Nathalie Lalevee ^1,2,7

^1. 

Equipe 5, C2VN INSERM 1263, Marseille, France

^2. 

Institut MarMaRa, Aix-Marseille Université, Marseille, France

^3. 

CNRS UMR 7288, IBDM, Marseille, France

^4. 

SMARTc, Marseille Protéomique—CRCM, Marseille, France

^5. 

Tagc, UMR 1090, Marseille, France

^6. 

Unit of Heart Failure and Valvular Heart Diseases, Department of Cardiology, North Hospital, Hôpitaux Universitaires de Marseille—AP-HM, Marseille, France

^7. 

Mediterranean Center of Cardio-Oncology, Aix-Marseille Université, Marseille, France

Introduction: Immunotherapy improves cancer outcomes but can cause myocarditis, with a high mortality risk. Immune checkpoint inhibitor-induced myocarditis (ICI-M) involves T-cell and macrophage infiltration in the myocardium, leading to tissue necrosis. The NLRP3 inflammasome pathway, triggered by IFN-γ, is overexpressed in ICI-M patients.

Objectives: To study ICI-M pathophysiology using a preclinical model to investigate electrophysiological disorders linked to myocardial immune infiltration and to explore molecular mechanisms in an IPSC cellular model from ICI-M patients.

Method: The preclinical ICI-M model was performed by injecting murine melanocytic cells into BALB/c mice treated with anti-PD1/anti-CTLA-4 therapy. IPSC-derived cardiomyocytes (CMs) and endothelial cells (ECs) from ICI-M patients and healthy donors were exposed to IFN-γ.

Results: A decrease in tumor volume (p0.01) was observed in mice with ECG disturbances, showing reduced QRS (p0.01) and T wave (p0.01) amplitudes, as well as a shorter repolarization time (p0.05). Impaired cardiac function correlated with increased CD3 transcript (p0.01) and PD-L1 overexpression in the myocardium. In hiPSC-derived CMs and ECs, IFN-γ (10 ng/mL, 72 h) upregulated immune-response genes (PD-L1, MHC-II, p0.001) and NLRP3 inflammasome genes (GBP5, GBP6, NLRC5, p0.0001). More dysregulated genes were found in ECs (FDR 0%, 2210 vs. 789 in CMs) after treatment. These genes are linked to the immune response, apoptosis, and inflammasome activation. HiPSC-ECs showed a decreased expression of cell communication genes and an increased expression of JAK-STAT/MAP kinases, NF-κB, and cytokine production genes. The inflammasome activated by IFN-γ differed between the cell types, with NLRP3 being activated in the CMs and AIM2 in the ECs. HiPSC-ECs from the ICI-M patients showed specific immune-response regulations, including PD-L2, TLR2 overexpression, and proinflammatory cytokines (CCL2, CCL5, IL-1β), linked to NLRP3 inflammasome activation.

Conclusions: The mice treated with ICIs showed ECG disturbances linked to myocardial inflammation. Our model highlights the key role of endothelial cells in the IFN-γ response and NLRP3 inflammasome regulation in ICI-M patient cells.

2.3. Comparative Analyses of Immunotoxins’ Efficacy on 2D and 3D Models of Sarcoma

Giulia Calafato ¹, Massimo Bortolotti ², Letizia Polito ², Andrea Bolognesi ²

^1. 

IRCCS Azienda Ospedaliero-Universitaria di Bologna, 40138 Bologna, Italy;

^2. 

Department of Medical and Surgical Sciences—DIMEC, Alma Mater Studiorum, University of Bologna, 40126 Bologna, Italy

Introduction: Sarcomas are mesenchymal tumors often associated with therapeutical resistance and recurrence. Immunotoxins (ITs) represent a promising strategy, joining the specificity of carriers to the cytotoxicity of plant toxins (ribosome-inactivating protein, RIP). Three-dimensional models represent a ground-breaking advance in cancer therapy, especially for predictive, high-throughput drug screening. Our study aimed to comparatively evaluate three ITs’ efficacy against TfR1-, EGFR1-, and Her2-expressing sarcoma cells, in 2D (adherent cells) and 3D (spheroids) models.

Methods: Three different ITs were obtained by chemical conjugation, firstly of the RIP saporin with Transferrin (Tf-IT), secondly of the RIP Ocymoidine with α-EGFR1 (α-EGFR1-IT), and thirdly of the RIP Ocymoidine with α-Her2 (α-Her2-IT). The ITs’ efficacy was evaluated on rhabdomyosarcoma (RD18) and osteosarcoma (U2OS) in dose–response viability experiments (an MTS colorimetric assay for the 2D model and an ATP luminometric assay for the 3D model) and using a Caspase 3/7 activation assay.

Results: In the RD18 and U2OS 2D and 3D models, the RIPs’ cytotoxicity was significantly enhanced after chemical conjugation with the carriers. The ITs strongly reduced cell viability, showing EC50 values of 1.3-2.3 logs lower than their corresponding unconjugated RIPs and carriers. Despite the increased complexity and penetration difficulty characterizing 3D models, the ITs were also effective in the 3D models, showing similar EC50 values to those of the 2D models. All the ITs were able to strongly activate Caspase 3/7.

Conclusions: Our data showed the strong efficacy of all the tested ITs. The strategy of targeting different antigens on tumor cells’ surfaces may also allow us to kill clones expressing mutated target antigens. RIPs trigger different cell death mechanisms, avoiding the selection of drug-resistant neoplastic clones. Thus, our results support the potential use of ITs as a therapeutic strategy in the treatment of sarcomas. Further experiments will be necessary to evaluate “in vivo” the tolerable doses of ITs and to compare their efficacy with the drugs approved by the FDA for sarcoma therapy.

2.4. Transcriptional Impact of Proinflammatory Cytokines in hiPSC-Derived Endothelial Cells and Cardiomyocytes: Insights into ICI-Induced Myocarditis

Conte Samantha ^1,2, Firoaguer Isaure ^1,2, Lledo Simon ^1,2, Tran Thi Tom ^1,2, El Yazidi Claire ^2,3, Chevillard Christophe ^2,4, Guieu Régis ^1,5, Puthier Denis ^2,4, Fenouillet Emmanuel ^1,2, Thuny Franck ^1,2,6, Cautela Jenifer ^1,2,6, Lalevée Nathalie ^1,2

^1. 

Aix Marseille University, INSERM, INRAE, C2VN, Marseille, France

^2. 

Aix Marseille University, MarMaRa Institute, Marseille, France

^3. 

Aix Marseille University, INSERM, MMG, Marseille, France

^4. 

Aix Marseille University, INSERM, TAGC, Marseille, France

^5. 

Laboratory of Biochemistry, Timone Hospital, Marseille

^6. 

Aix Marseille University, AP-HM, North Hospital, Department of Cardiology, Medi-CO Center, Marseille, France

Introduction: Inflammatory cytokines, particularly IFN-γ, are highly elevated in biopsies and the peripheral blood of patients with ICI-induced myocarditis (ICI-M). While the NLRP3 inflammasome has been implicated in ICI-M pathophysiology, its underlying mechanisms remain poorly understood.

Objective: This study investigates the transcriptional effects of inflammatory cytokines associated with ICI-M on endothelial cells (hiPSC-ECs) and cardiomyocytes (hiPSC-CMs) derived from a healthy individual’s hiPSC clone.

Method: The transcriptional profiles of both the hiPSC-derived endothelial cells (hiPSC-ECs) and cardiomyocytes (hiPSC-CMs) were analyzed following the IFN-γ treatment. Inflammatory responses, cell mortality, and apoptosis were assessed after stimulation with proinflammatory molecules, either through individual IFN-γ exposure or a combined cytokine cocktail (CCL5, GZMB, IL-1β, IL-2, IL-6, IFN-γ, and TNF-α), to characterize their effects on cellular function and viability better.

Results: Our findings reveal that hiPSC-ECs are highly sensitive to cytokine treatment, exhibiting significant mortality and marked transcriptomic changes in immunity- and inflammation-related pathways. In contrast, hiPSC-CMs show lower transcriptomic alterations, reduced mortality, and less apoptosis. In both cell types, cytokine treatment upregulates key components of the NLRP3 inflammasome pathway, including regulators (GBP5, GBP6, P2X7, NLRC5), a core component (AIM2), and an effector (GSDMD). Notably, cytokine treatment increases the GBP5 protein expression and CASP-1 cleavage in hiPSC-CMs, mirroring observations in endomyocardial biopsies from ICI-M patients.

Conclusions: This cellular model provides valuable insights into endocardial and microvascular dysfunctions in inflammatory conditions and their impacts on cardiomyocytes. These findings establish a robust experimental model for investigating the response of hiPSC-derived cardiac cells to the inflammatory cytokines implicated in immuno-mediated myocarditis. Moreover, they provide essential reference values for comparative studies using hiPSC clones derived from affected patients, thereby facilitating a deeper understanding of the disease mechanisms.

3. Tumor Microenvironment

3.1. Comprehensive Transcriptomic Profiling and ceRNA Network Analysis Reveal Hub Genes and Therapeutic Targets in HPV-Positive Head and Neck Squamous-Cell Carcinoma

Kunpeng Wang ¹, Boyu Pan ², Rui Li ²

^1. 

OMFS Department, Uygur Medical Hospital of Hetian Prefecture, Xinjiang Uygur Autonomous Region, Hetian 848000, China

^2. 

Oncological Department, Uygur Medical Hospital of Hetian Prefecture, Xinjiang Uygur Autonomous Region, Hetian 848000, China

Objective: Human papillomavirus (HPV) infection is a significant risk factor for head and neck squamous-cell carcinoma (HNSCC). This study aims to identify specific signaling pathways, biomarkers, and potential targeted drugs for HPV-positive HNSCC through a competitive endogenous RNA (ceRNA) network analysis.

Methods: High-throughput transcriptomic sequencing was performed on UPCI:SCC154 (HPV-positive HNSCC), Cal27 (HPV-negative HNSCC), and normal human oral keratinocytes to identify differentially expressed mRNAs and miRNAs associated with HPV-positive HNSCC. The interactions of competitive endogenous RNAs were predicted using TargetScan and miRanda, and protein–protein interactions were predicted using the String database to construct a ceRNA network. A network topology analysis was employed to identify hub genes in HPV-positive HNSCC. Further, upstream miRNAs of the hub genes were screened by integrating multiple databases, including encori, mircode, miRDB, miRTarBase, miRWalk, and picTAR, to construct miRNA-mRNA signaling axes. A drug repositioning analysis was conducted using the Connectivity Map database to predict small-molecule drugs targeting the hub genes. Molecular docking and dynamics simulations were used to evaluate the binding affinity and stability of the small molecules with the target proteins. The efficacy of the drugs was validated through cellular functional assays.

Results: A total of 3,253 differentially expressed mRNAs and 391 miRNAs were identified in HPV-positive HNSCC, and RPAP2 was identified as a hub gene. The integrated multi-database analysis revealed that hsa-miR-181b-3p potentially targets and regulates RPAP2. Drug repositioning screening identified five potential drugs, among which molecular docking and dynamics simulations indicated that Enzastaurin exhibited excellent binding affinity and stability with RPAP2. Cellular experiments demonstrated that, compared to cisplatin, Enzastaurin significantly reduced the viability of HPV-positive HNSCC cells and suppressed the expression levels of RPAP2 and hsa-miR-181b-3p.

Conclusions: The miR-181b-3p/RPAP2 axis plays a key regulatory role in HPV-positive HNSCC. Enzastaurin demonstrates promising therapeutic potential, offering a foundation for targeted drug development.

3.2. DAVID Analysis Suggests Novel Chemoresistance Pathways in Ovarian Cancer

Aditya Vayalapalli, Ilana Chefetz

• Biomedical Sciences, School of Medicine, Mercer University, Macon, 31207, Georgia, USA

Ovarian cancer is the deadliest gynecological malignancy, with around 310,000 women worldwide being diagnosed with the cancer in 2020. Resistance to chemotherapeutics is inevitable in most ovarian cancer patients, but little is known about the precise mechanisms of chemoresistance in ovarian cancer. This study aimed to uncover novel chemoresistance mechanisms using the bioinformatics tools GEO2R and DAVID by compiling a list of differentially expressed genes and then clustering them into specific expression patterns. Differential gene expression data of in vitro studies comparing paclitaxel- and cisplatin-resistant subtypes to wild-type counterparts were obtained from GSE73935 and GSE26465 and analyzed in GEO2R. A common list of 8697 differentially expressed genes between both datasets was extracted using the dplyr package in R. The list of common genes was input into the DAVID functional annotation clustering tool. The results were considered statistically significant at the threshold of (p0.05) and a false discovery rate under 0.05. The protein–protein interactions network was used in order to ascertain the molecular mechanisms of specific differentially expressed proteins in relation to the clusters they relate to. Genes that were differentially expressed fell into established patterns of chemoresistance that well established in ovarian cancer, such as epithelial–mesenchymal transition and autophagy. However, expression profiles consistent with chemoresistance pathways in other cancers, but not well established in ovarian cancer, were also identified, namely pathways of transcription regulation, ubiquitin-binding, protein transport, the TGF-beta signaling pathway, cell projection, and cell polarity. These findings suggest novel chemoresistance pathways that could be targeted to resensitize chemoresistant cells to chemotherapeutics or for the early prediction of chemoresistance in patients.

3.3. Alterations in the Expression of a Group of Genes Associated with the Epithelial–Mesenchymal Transition in Lung Cancer: A Pilot Study

Irina Valerievna Pronina

• Emanuel Institute of Biochemical Physics, Russian Academy of Science, Moscow, Russia

A large number of diagnosed cases of lung cancer occur in late stages and are accompanied by metastasis, which makes research into the epithelial-to-mesenchymal transition (EMT) genes relevant. The genes responsible for the EMT include E-cadherin (CDH1), which is involved in the formation of tight junctions between epithelial cells, soluble growth factors, and some transcription factors (e.g., ZEB1, ZEB2).

The aim of this study was to evaluate the expression alterations in BMI1, CDH1, SNAI1, SNAI2, ZEB1, and ZEB2 in tumors compared to histologically normal lung tissues.

The study used 50 paired RNA samples with RIN ≥ 7 isolated from tumors, including 37 non-metastatic and 13 metastatic tumors, and normal lung tissues. The expression levels were assessed by RT-qPCR. B2M and ACTB were used as reference genes. The statistical processing of the results was performed by ANOVA using the CFX Maestro software (Bio-Rad, Hercules, CA, USA).

The expression of six genes associated with the EMT—BMI1, CDH1, SNAI1, SNAI2, ZEB1, and ZEB2—was analyzed. CDH1 expression increased by 3.63 times in the tumors compared to the norm, p ≤ 0.05; moreover, in the non-metastatic tumors, it increased by 3.47 times, but in the metastatic tumors, it increased by 4.21 times, p ≤ 0.05. ZEB1 and ZEB2 expression decreased by 2.32 and 2.01 times, p ≤ 0.05, and their expression in the metastatic tumors decreased by 3.22 and 3.01 times, p ≤ 0.05. The expression levels of BMI1, SNAI1, and SNAI2 changed by a factor of −1.51 to 1.53; however, these data were not statistically significant.

Based on the results of this study, we can conclude that E-cadherin in lung cancer promotes either the epithelial-to-mesenchymal or mesenchymal-to-epithelial transition and, consequently, metastasis. We also showed a suppression of the expression of transcription factors encoded by ZEB1 and ZEB2. The role of BMI1, SNAI1, and SNAI2 in lung cancer is likely limited, as their expression remained largely unchanged.

3.4. Can Statins Transform Human Colon Cell Behavior? Unveiling the Insights from Raman Imaging and AFM

Karolina Beton-Mysur, Beata Brożek-Płuska

• Institute of Applied Radiation Chemistry, Faculty of Chemistry, Lodz University of Technology, Wroblewskiego 15 93-590 Lodz, Poland

Over time, advancements in cancer diagnostics and therapies have been remarkable. This study compares normal (CCD-18Co) and cancerous (CaCo-2) cell lines from the human gastrointestinal tract, analyzing their nanomechanical and biochemical properties to identify potential cancer biomarkers that could enhance oncological diagnostics. The research examines the effects of 24 and 48 h of 10 µM supplementation with statins—mevastatin, lovastatin, and simvastatin—on the biochemical and nanomechanical properties of these cells, employing Raman imaging and AFM techniques. The innovative integration of these techniques provides a comprehensive biochemical and nanomechanical characterization of normal, cancerous, and statin-treated cancer cells, highlighting how different statins influence cellular properties.

A primary objective of this study was to identify statistically significant differences between normal and cancerous colon cells, including cancer cells treated with mevastatin, simvastatin, and lovastatin, based on their vibrational features. We systematically explored how Raman imaging and spectroscopy respond to normal and cancerous human cells, both untreated and treated with statins. Our approach demonstrates a fast, cost-effective method for visualizing cellular structures and performing virtual staining. By incorporating Raman intensity into pseudo-color images, these high-resolution, label-free images allow for the direct analysis of cellular substructures, facilitating the detection of biochemical changes characteristic of cancer progression and aiding in the assessment of anticancer therapies.

Using Raman data, we analyzed and compared the vibrational features of normal, cancerous, and statin-treated cancer cells. Average spectra and Raman band intensity ratios for key biological components—proteins, nucleic acids, and lipids—enabled detailed insights into the molecular changes associated with cancer and its treatment.

Our findings provide evidence supporting the hypothesis that the long-term use of lipophilic statins may influence cancer incidence, including specific cancer types. Additionally, statins may enhance chemotherapy sensitivity and improve clinical outcomes for patients already diagnosed with cancer.

3.5. Quantum Artificial Intelligence in Tumor Classification: An Innovative Method for Biomedical Data Analysis

Emine Akpinar

• Department of Physics, Graduate School of Science and Engineering, Davutpasa Campus, Yıldız Technical University, Istanbul, 34220, Turkey

Abnormal cell growth in the brain characterizes brain tumors, which are primarily diagnosed through histopathological examination. Noninvasive neuroimaging techniques, such as MRI, provide critical diagnostic insights; however, the size and complexity of MRI data pose challenges for effective analysis. While traditional AI methods are effective in medical data analysis, factors such as the expansion of high-resolution medical datasets and noise levels in the data impact the diagnostic process. Recent studies have shown that the application of quantum AI technologies in healthcare not only addresses these issues but also accelerates complex data analyses, providing a significant advantage, especially in dealing with heterogeneous and unevenly distributed datasets. In this study, a quantum deep neural network (QDNN) model is proposed to distinguish four different classes—glioma, meningioma, pituitary tumors, and cases without tumors—based on data from 7023 individuals obtained from the Kaggle open data portal, aiming for the highest accuracy. During preprocessing, image enhancement techniques were applied using the OpenCV library to optimize the data quality. Subsequently, the proposed QDNN model was employed for classification. In the model, amplitude encoding was utilized to map MR image data from a classical space to a quantum Hilbert space, followed by a multi-layer parameterized quantum circuit (multi-layer PQC). The multi-layer PQC model consists of single-qubit Rx gates, along with a CNOT gate that provides circular entanglement between qubits. As a result, the proposed model achieved final training loss and validation loss values of 0.61 and 0.64, respectively, while the training accuracy and validation accuracy values were 0.76 and 0.77, respectively. When compared to a classical DNN model with a similar number of parameters, the proposed quantum model demonstrates superior performance in terms of both accuracy and the total processing time. These results highlight the potential of quantum AI in improving diagnostic accuracy in biomedical imaging.

3.6. Translational Clinical Feasibility of Recent Advances in Liquid Biopsy Technology in Effusion Fluids: Preliminary Results

Domenico Damiani ¹, Christine Mian ¹, Christine Schwienbacher ¹, Ilaria Girolami ¹, Esther Hanspeter ¹, Giovanni Negri ¹, Georgia Levidou ²

^1. 

Department of Pathology, Provincial Hospital of Bolzano (SABES-ASDAA), Lehrkrankenhaus der Paracelsus Medizinischen Privatuniversität, via Lorenz Böhler, 5, 39100 Bolzano-Bozen, Italy

^2. 

Department of Pathology, Nuremberg Clinic, Paracelsus Medical University, 90419 Nuremberg, Germany

Introduction: Recent advancements in molecular profiling technologies based on liquid media (so-called liquid biopsy) have achieved a significant increase in detection sensitivity, enhancing our power of investigation for biofluids and suggesting their potential integration into pivotal diagnostic and predictive molecular analyses, such as for non-small-cell lung cancer. Pre-analytical variables play a crucial role in the accuracy of analysis results. Evidence suggests that supernatant fluid obtained from cytological sample centrifugation is enriched with nucleic acids and less susceptible to nucleic acid degradation during processing compared to cytoblock samples. This study reports preliminary results from a recent workflow implementation of a liquid biopsy in a routine molecular biology laboratory within the context of a validation study.

Methods: Eleven cytological samples of lung adenocarcinoma and corresponding cytoblock slides underwent a morphological analysis and review in terms of cellularity, cell integrity, necrosis/debris, and the presence of artifacts. Following the NSG analysis of both the effusion fluid and the cytoblock, a data analysis and a comparison of the results were performed, focusing on concordance and the pre-analytical and analytical variables observed in the two groups.

Results: Molecular profiling led to the detection of an actionable alteration in all the patients, with more robust results, evaluated in terms of the variant allele frequency (VAF) in the supernatant group (53.3%) in comparison with the cytoblock group (26.49%). This study resulted in a 100% concordance in molecular profiling between the cytoblock and supernatant analyses, with the latter showing higher yields in nucleic acid extraction, suggesting the possibility of achieving clinically useful results for running molecular profiling on biofluid materials.

Conclusions: Liquid biopsy technology has gained enough robustness to be a useful tool in routine analysis. Biofluid application in tumor molecular profiling is a promising field of exploration with outstanding potential in the case of insufficient cytologic material, contributing to improved patient management and, thus, avoiding repetitive procedures and optimizing the overall efficiency and cost-effectiveness of diagnostic practices.

3.7. Unveiling Natural Inhibitors for ABCC1 (MRP1) Membrane Transporter Through Molecular Docking and Molecular Dynamics Simulations

Roya Rahimivaghar ^1,2, Seyed Shahriar Arab ¹, Justin Tze-Yang Ng ², Yaw Sing Tan ²

^1. 

Department of Biophysics, Faculty of Biological Sciences, Tarbiat Modares University, Tehran, Iran

^2. 

Bioinformatics Institute, Agency for Science, Technology and Research (A*STAR), 30 Biopolis Street, #07-01 Matrix, Singapore 138671, Singapore

ABC transporters are fascinating proteins responsible for transporting a variety of substrates through the hydrolysis of ATP. Some ABC transporters are multidrug-resistant (MDR), a trait commonly associated with human cancers and pathogenic microbes. Their ability to transport toxic substances and drugs across membranes, even against the concentration gradient, leads to a reduced concentration of drugs inside cells, which diminishes the drug’s effectiveness. Natural compounds, such as polyphenols and flavonoids, have anticancer properties. By inhibiting ATP hydrolysis, they can potentially inhibit MDRs in cancer cells, reduce the activity of these proteins, and enhance the therapeutic effects of anticancer drugs. In this study, we investigate the inhibitory effects of 77 of these compounds on the nucleotide-binding domains (NBDs) of ABCC1 (MRP1) by using molecular docking and molecular dynamics (MD) simulation. The results indicate that five compounds, namely (-)-catechingallat, limonin, naringin, rhoifolin, and robinin, had high binding affinities, with values of −7.8, −8.5, −8.3, −8.3, and −8.5 in NBD1 and −7.1, −7.9, −8.2, −7.9, and −7.7 in NBD2, respectively; in comparison, ATP showed binding affinities of −6.8 and −7.1 in NBD1 and NBD 2, respectively; all the compounds occupied the same binding sites as ATP, namely Asp793 and Tyr831 in NBD1 and Arg1445 in NBD2. A molecular dynamics trajectory analysis of the NBDs and ligands revealed these domains were stable throughout 200 ns MD simulations. The MD simulations confirm the stability of the complex formed by the interaction of five ligands with NBD, characterized by structural compactness and minimal to no fluctuations. This in silico study offers key information for developing potential ATP inhibitors, suggesting that NBDs could be suitable binding sites for the flavonoid family. The discovery of novel MDR-inhibiting compounds has the potential to make cancer treatment more effective for all types of cancers, making it a comprehensive solution to drug resistance.

4. Immune System, Tumor Immunology, and Autoimmune Disease

4.1. Efficacy and Safety of Tocilizumab in the Treatment of Rheumatoid Arthritis: An Umbrella Review

David Silva ¹, Fernando Moreira ², Marlene Santos ²

^1. 

Escola Superior de Saúde, Instituto Politécnico do Porto, Rua Dr. António Bernardino de Almeida, 4200-072, Porto, Portugal

^2. 

REQUIMTE/LAQV, Escola Superior de Saúde, Instituto Politécnico do Porto, Rua Dr. António Bernardino de Almeida, 4200-072, Porto, Portugal

Introduction: Tocilizumab (TCZ) is a humanized immunoglobulin G1 monoclonal antibody that targets and inhibits the interleukin-6 (IL-6) receptor. As a selective inhibitor of this cytokine, TCZ offers a targeted approach in managing inflammatory conditions. Given its specificity and potential therapeutic benefits, this article aims to provide a comprehensive summary of the efficacy and safety of TCZ in the treatment of RA.

Methods: This study adhered to the structure of an Umbrella Review, which synthesizes multiple systematic reviews and clinical studies to provide a broad understanding of the subject matter. The review was conducted between October 2023 and August 2024, with a focus on patients diagnosed with rheumatoid arthritis. A thorough search was performed using the PubMed and Cochrane databases to identify relevant studies. The inclusion criteria were based on systematic reviews, while exclusion was applied to studies based on the article type and the publication date.

Results: Seventeen studies were included in the analysis. These studies utilized the American College of Radiology (ACR) criteria to assess the efficacy of TCZ in comparison to other conventional and biologic DMARDs, as well as a placebo. The results revealed that TCZ monotherapy demonstrated superior efficacy in reducing disease activity and improving physical function when compared to alternative treatments. Safety was evaluated by reviewing adverse reactions and infections associated with TCZ. Overall, the incidence of adverse events was similar to that observed with other biologic therapies, suggesting that TCZ does not pose an increased risk in this regard.

Conclusions: Tocilizumab has proven to be an effective and safe option in the treatment of rheumatoid arthritis, offering distinct advantages in both efficacy and safety compared to other conventional and biological DMARDs. The findings from this review support its continued use, including as adjunctive therapy in managing RA, particularly for patients who do not respond adequately to traditional treatments.

4.2. Anti-Arthritic Potential of Artemisia Herba-Alba in Carrageenan- and Complete Freund’s Adjuvant-Induced Arthritis Models in Rats

Hicham Wahnou ¹, Zaynab Ouadghiri ², Martin Ndayambaje ², Salma Benayad ³, Youness Limami ³, Mounia Oudghiri ²

^1. 

Laboratory of Immunology and Biodiversity, Faculty of Sciences Ain Chock, Hassan II University, B.P 2693, Maarif, Casablanca, Morocco

^2. 

Laboratory of Immunology and Biodiversity, Faculty of Sciences Ain Chock, Hassan II University, B.P2693, Maarif, Casablanca, Morocco

^3. 

Laboratory of Health Sciences and Technologies, Higher Institute of Health Sciences, Hassan First University of Settat, Settat 26000, Morocco

Arthritis is a chronic inflammatory disorder characterized by progressive joint damage and immune dysregulation, significantly impacting patients’ quality of life. Conventional treatments, such as NSAIDs and corticosteroids, while effective, often cause adverse effects, such as gastrointestinal bleeding, cardiovascular risks, and osteoporosis, particularly with long-term use. This study explores the anti-arthritic potential of Artemisia herba-alba, a medicinal plant with traditional anti-inflammatory uses, in carrageenan- and complete Freund’s adjuvant (CFA)-induced arthritis models in rats.

Two arthritis models were used: (1) CFA-induced arthritis, where inflammation was triggered by means of a single subcutaneous CFA injection into the hind paw, evaluated over 15 days, and (2) carrageenan-induced arthritis, induced by means of repeated intra-articular carrageenan injections into the hind paw over 30 days. Male Wistar rats were divided into control, arthritic, and treatment groups. The treatment groups received oral A. herba-alba extracts (250 mg/kg or 500 mg/kg), while the positive control group received indomethacin (3 mg/kg). The hematological analysis quantified serum neutrophils and monocytes, and the histopathological examination assessed joint tissues using H&E staining.

A. herba-alba extracts demonstrated dose-dependent anti-inflammatory effects, with the 500 mg/kg dose outperforming indomethacin. The hematological analysis showed significant reductions in neutrophils and monocytes, indicating systemic immune modulation. The histopathological findings revealed reduced osteoclast activity, decreased neutrophil infiltration, and preserved synovial tissue integrity. The 500 mg/kg dose notably mitigated osteoclastic bone resorption and prevented synovial hyperplasia.

These results highlight Artemisia herba-alba as a potent natural anti-inflammatory agent with systemic and localized effects, offering a safer alternative for arthritis management. Its efficacy at 500 mg/kg, surpassing indomethacin, underscores its therapeutic potential. Further research, including clinical trials, is needed to confirm its safety and efficacy in humans and to elucidate its mechanisms of action.

4.3. Exploring the Advances in Plaque Psoriasis Research: A Scientometric Analysis and Visualization (2000–2025)

Parsa Dar ¹, Azam Mehmood Dar ², Cui Wenqing ³, Jun Ming Chen ¹, Hua Yu ^3,4

^1. 

Institute of Chinese Medical Sciences, State Key Laboratory of Quality Research in Chinese Medicine, University of Macau, Macao, China

^2. 

Department of Bio products and Biosystems Aalto University, Puunjalostustekniikka 1, Vuorimiehentie 1, 02150 Espoo Finland

^3. 

State Key Laboratory of Quality Research in Chinese Medicine, Institute of Chinese Medical Sciences, University of Macau, Macao SAR, China

^4. 

Macao Centre for Research and Development in Chinese Medicine, Institute of Chinese Medical Sciences, University of Macau, Macao SAR, China

Despite numerous studies investigating plaque psoriasis, there exists a paucity of research regarding scientometric analysis. This document presents a scientific analysis and historical overview of plaque psoriasis research. This study employs the prior methodology to analyze plaque psoriasis research from 2000 to 2025. It identifies hotspots, patterns of plaque psoriasis, and keyword trends. A scholarometric analysis identified the primary research domains of plaque psoriasis: field distribution, knowledge structure, subject evolution, and topic emergence. The induction factor, comorbidity, etiology, therapy, and animal models illustrate the incidence, progression, and treatment of depression. A scientometric analysis identified 13,646 documents on plaque psoriasis over a span of 25 years. These results establish a robust basis for subsequent research. We identified 13,646 articles on plaque psoriasis. Multiple journals addressed plaque psoriasis, including the British Journal of Dermatology, the American Academy Journal, and the European Academy Journal. Manchester University was the most productive, with 399 publications. Research on psoriasis was most abundant in the United States, yielding 4141 publications across dermatology, pharmacology, rheumatology, and health care. Comorbidities, interleukin-23, drug survival, nail and plaque psoriasis, myocardial infarction, weight loss, diabetes, obesity, metabolic syndrome, cyclosporine, depression, IL-17, IL-23, adherence, and arthritis seem to be interconnected.

The citation analysis delineated the foremost researchers, institutions, and publications in plaque psoriasis, offering a thorough overview. The increase in publications indicates that this topic has attracted academic interest. The results underscore the significance of collaboration, interdisciplinary approaches, and resource allocation in marginalized sectors for researchers, funding organizations, and governmental entities. This study advocates for the utilization of traditional Chinese medicine and herbal remedies in the treatment of plaque psoriasis, citing their reduced side effects.

4.4. Histological and Immunohistochemical Study of Anti-Inflammatory Effect of Inula viscosa (L). Aiton Leaf Extract in Formaldehyde-Induced Arthritis in Mice

Sara Ouari ¹, Nadia Benzidane ², Chahla Bencharif ³, Noureddine Bribi ⁴

^1. 

Laboratory of Applied Biochemistry, Faculty of Nature and Life Science, University Ferhat Abbes Setif 1, 19000, Setif, Algeria

^2. 

Laboratory of Applied Biochemistry, Faculty of Nature and Life Science, Setif 1 Ferhat Abbas University, 19000, Setif, Algeria

^3. 

Pathology Laboratory, Khelil Amrane University Hospital of Bejaia, 06000, Bejaia, Algeria

^4. 

Laboratory of Plant Biotechnology and Ethnobotany, Faculty of Nature and Life Sciences, University of Bejaia, Bejaia, 06000, Algeria

Introduction: Rheumatoid arthritis (RA) is a chronic and complex inflammatory disorder characterized by the inflammation of the synovial membrane (synovitis), leading to cartilage and bone damage in articular joints. Given the adverse effects and different and non-affective responses of patients to synthetic, disease-modifying, anti-rheumatic drugs (DMARDs) and nonsteroidal anti-inflammatory drugs (NSAIDs), it is of interest to use medicinal plants exhibiting encouraging therapeutic results and fewer side effects. Objective: This study aimed to investigate the anti-arthritic effect of Inula viscosa (I. viscosa) on the progression of arthritis in an NMRI mouse model.

Methods: Mice were divided into six groups (n = 6) as follows: normal control, disease control, Diclofenac group (10 mg/kg, p.o. daily), and three groups who were treated daily with 50, 100, and 200 mg/kg of IVME (p.o.). Formaldehyde models were obtained by means of the sub-plantar administration of 20 µL of formaldehyde (3.75% v/v) into the right hind paws of NMRI albino mice on the first and third of the ten experimental days. Joint diameters were measured; the arthritis severity was evaluated by means of the inhibition of the paw oedema; histological changes, synovial hyperplasia, and immune cell infiltration were evaluated using histological and immunohistochemical analyses of CD3+, CD20+, and CD68+; and staining markers of ankle joint tissue sections were analyzed with the QuPath v0.5.1 software tool using adaptive thresholding to quantify the percentage staining of positive cells.

Results: The administration of I. visocsa (at a low dose of 50 mg/kg) significantly (*** p < 0.001) ameliorated the induced arthritis severity, resulting in the restoration of the paw diameter, (* p < 0.05), reduced hyperplasia of the synovial membrane, and bone erosion, and significantly (* p < 0.1) decreased CD68+-staining immune cell infiltration.

Conclusions: These findings suggest that I. viscosa leaves have an anti-arthritic property, which is due to their anti-inflammatory effect, probably through cytokine regulation.

4.5. Natural Extracts and Autoimmunity: The Therapeutic Potential of Kiwi Polyphenols

Franklin Chamorro ¹, Rafael Nogueira-Marques ², A. G Pereira ³, J. Echave ¹, A.O.S. Jorge ^1,4, Ezgi Nur Yuksek ¹, M.A. Prieto ¹

^1. 

Universidade de Vigo, Nutrition and Bromatology Group, Department of Analytical Chemistry and Food Science, Instituto de Agroecoloxía e Alimentación (IAA)—CITEXVI, 36310 Vigo, Spain

^2. 

Universidade de Vigo, Nutrition and Bromatology Group, Department of Analytical Chemistry and Food Science, Instituto de Agroecoloxía e Alimentación (IAA)—CITEXVI, 36310 Vigo, Spain

^3. 

Investigaciones Agroalimentarias Research Group, Galicia Sur Health Research Institute (IIS Galicia Sur). SERGAS-UVIGO, Vigo, Spain

^4. 

REQUIMTE/LAQV, Instituto Superior de Engenharia do Porto, Instituto Politécnico do Porto, Rua Dr. António Bernardino de Almeida 431, 4249-015 Porto, Portugal

Autoimmune diseases are characterized by an exacerbated immune response against the body’s own tissues. The main pathologies include rheumatoid arthritis, systemic lupus erythematosus, multiple sclerosis, psoriasis, and inflammatory bowel disease. All these pathologies are based on oxidative stress and chronic inflammation. Therefore, the search for alternative therapies based on natural compounds with antioxidant, anti-inflammatory, and immunomodulatory properties has become highly relevant in biomedical research. In this sense, polyphenols extracted from the skin of green kiwi (Actinidia deliciosa) have shown beneficial effects in the regulation of the immune system. This review aims to describe how polyphenols extracted from the skin of green kiwi interact with key enzymes and proteins in the human body to reach their therapeutic target. In this sense, kiwi polyphenols can exert positive effects on autoimmune diseases due to their ability to modulate proinflammatory cytokines, including tumor necrosis factor alpha (TNF-α), interleukin-6 (IL-6), and interleukin-1β (IL-1β). On the other hand, kiwi polyphenols can act as potent scavengers of reactive oxygen species (ROS), reducing oxidative damage at the cellular level and protecting DNA, lipids, and proteins. Finally, green kiwi polyphenols have the ability to modulate the adaptive immune response, influencing the differentiation and activation of T lymphocytes, decreasing the activation of Th1 and Th17 cells, which are involved in the chronic inflammation of various autoimmune diseases. Preclinical studies provide evidence on the beneficial effects of polyphenols extracted from kiwifruit peel in autoimmune diseases; however, clinical studies in humans are needed to determine their efficacy, safety, and possible therapeutic applications. The findings of this review contribute to the elucidation of the biochemical interactions of polyphenols extracted from green kiwifruit in autoimmune pathologies, highlighting the opportunities and limitations of their use as a natural strategy for the treatment of autoimmune diseases.

4.6. Pulmonary Fibrosis and Lung Cancer: Unity of and Differences in Cellular Processes

Olga Pershina ¹, Angelina Pakhomova ^1,2, Olga Rybalkina ³, Ildar Uzyanbaev ^1,4, Lena Kogai ¹, Lubov Sandrikina ^1,2, Elena Fedorova ¹, Valentina Sedinova ¹, Vadim Zhdanov ¹

^1. 

Laboratory of Regenerative Pharmacology, Goldberg ED Research Institute of Pharmacology and Regenerative Medicine, Tomsk National Research Medical Centre of the Russian Academy of Sciences, Tomsk, Russia

^2. 

Normal Physiology Department, Siberian State Medical University, Tomsk, Russia

^3. 

Goldberg ED Research Institute of Pharmacology and Regenerative Medicine, Tomsk National Research Medical Centre of the Russian Academy of Sciences, Tomsk, Russia

^4. 

Siberian State Medical University, Tomsk, Russia

Despite the multiple identified common genetic, molecular, and cellular processes connecting pulmonary fibrosis (PF) and lung cancer (LC), there is still no clear answer about the contribution of each of them to the development of pathology. It is known that most cell populations involved in the development of PF are involved in LC. At the same time, there are differences that can be explained by the specificity of PF development or changes specific to LC. We evaluated changes in the lung and blood during the modeling of PF, LC, and PF+LC. To induce PF in male C57BL6 mice, we used bleomycin; on the 15th day after bleomycin administration, we modeled non-small-cell LC by introducing Lewis lung carcinoma (LLC) cells. The material for this study was collected on the 28th day of the experiment and was analyzed by histology and cytometry. In the lung tissue of mice, against the background of PF and LC modeling, an increase in pathological changes in the lungs was observed compared to each pathological situation separately. Disorders of the lungs in the group of animals with PF and LC were based on the increased migration of populations of CD309+ endothelial cells from the blood to the lungs, an increase in atypical/hybrid cells (CD45+CD326+), and cancer stem cell markers CD90, CD117, CD274, CD276, and EGFR in various combinations in the lung tissue. Biomarkers confirming tumor development against the background of pulmonary fibrosis are populations of cancer stem cells, atypical/hybrid cells, endothelial cells, hematopoietic stem cells, myeloid fibrocytes, and changes in the composition of CD4+ T-cells. Further clinical and experimental studies are needed to determine the role of these changes and to find biomarkers that predict the progression of pulmonary fibrosis and the risk of developing lung cancer.

4.7. Recent Advances in Nanomedicine for Skin Cancer Immunotherapy: Focus on Nanosystems, Drug Repurposing, and Co-Loading for Synergistic Effects

Patrícia C. Pires

• Faculty of Pharmacy (FFUC-UC), University of Coimbra, and Health Sciences Research Centre, University of Beira Interior (CICS-UBI), Portugal

In skin cancer treatment, the often-severe side effects and limited efficacy of immunotherapies have prompted the development of novel therapeutic strategies. Hence, innovative formulations using nanosystems have emerged, which, due to having a nanometric size, being able to encapsulate and protect loaded drug molecules, leading to controlled drug release, providing increased permeation, and having tunable properties, have proven to improve therapeutic outcomes. Among the many types of nanoplatforms, nanoemulsions and nanoemugels have been some of the most successfully developed for topical immunomodulatory skin cancer treatment, including nanoemulgels, combining the immunomodulatory effects of imiquimod and the anti-inflammatory properties of curcumin, and nanoemulsions containing plant-derived anticarcinogenic and immunosuppressive molecule triptolide, which have led to increased skin drug permeation and retention and low toxicity in tissue models. On the other hand, drug repurposing, making use of already-marketed drug molecules and repositioning them for antitumor effects, has also been described for cancer immunotherapy, with doxycycline, a broad-spectrum antibiotic with recognized immunosuppressive and anticancer properties, having been incorporated into multifunctional hybrid electrospun nanofibrous scaffolds containing hydroxyapatite nanoparticles, yielding synergistic anticancer effects in in vitro skin cancer models. Furthermore, dual drug-loaded nanosystems have also proven to be beneficial in tumor immunotherapy, including immunoliposomes, simultaneously encapsulating the chemotherapeutic agent 5-fluorouracil and the antibody cetuximab, or immunoliposomes co-loaded with the immunomodulating compound bufalin and the anti-CD40 antibody, revealing increased and synergistic anticancer effects in in vitro and in vivo models of skin cancer, with or without the added use of iontophoresis, resulting in increased cellular uptake via endocytosis mediated by antigen–antibody interaction, simultaneous and long-lasting antigen delivery, with the induction of tumor apoptosis, and decreased tumor volume and tumor weight. Therefore, the use of nanosystems, drug repurposing, and synergistic dual drug-loading might be key to ensuring the success of future skin cancer immunotherapies.

4.8. Shared Genetic Risks and Pathways Between Asthma and Celiac Disease

Garnet Eister ¹, Hui-Qi Qu ¹ and Hakon Hakonarson ^1,2,3,4

^1. 

The Center for Applied Genomics, Children’s Hospital of Philadelphia, Philadelphia, PA 19104, USA

^2. 

Department of Pediatrics, The Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA 19104, USA

^3. 

Division of Human Genetics, Children’s Hospital of Philadelphia, Philadelphia, PA 19104, USA

^4. 

Division of Pulmonary Medicine, Children’s Hospital of Philadelphia, Philadelphia, PA 19104, USA

^5. 

Faculty of Medicine, University of Iceland, 101 Reykjavik, Iceland

Introduction: Asthma has a strong allergic component, while Celiac Disease (CD) is an autoimmune disorder triggered by gluten ingestion in genetically predisposed individuals. This study investigates whether asthma and CD share genetic risk factors, offering insights into their biological interplay and overlapping pathways.

Methods: Pediatric patients diagnosed with asthma (n = 4848) or CD (n = 346) were compared to controls who had never been diagnosed with any allergic or autoimmune diseases (n = 1963). Ninety-eight patients were diagnosed with both diseases. Asthma (PGS002727) and CD (PGS002067) polygenic risk scores (PRSs) were analyzed separately for females and males. All the participants were of European ancestry and recruited by the Center for Applied Genomics (CAG) at the Children’s Hospital of Philadelphia (CHOP).

Results: We conducted a cross-examination, excluding any patients diagnosed with both asthma and CD to avoid potential confounding effects. The key findings are as follows:

• Female Asthma Patients: Increased asthma PRS (p = 2.37 × 10⁻⁵) and CD PRS (p = 0.018).
• Female CD Patients: Elevated asthma PRS (p = 0.007) and CD PRS (p = 1.04 × 10⁻²⁵).
• Male Asthma Patients: Increased asthma PRS (p = 4.77 × 10⁻⁴); CD PRS not significant (p = 0.420).
• Male CD Patients: Higher asthma PRS (p = 9.83 × 10⁻⁶) and CD PRS (p = 4.31 × 10⁻¹⁶).
• Controls: Asthma and CD PRS correlated significantly (females: r = 0.249, p = 6.39 × 10⁻¹⁵; males: r = 0.232, p = 8.56 × 10⁻¹⁴).

Pathway PRS analyses using ImmuneSigDB C7 (5219 gene sets) showed 630 significantly altered pathway CD PRSs after Bonferroni correction. The pathway PRS associations for asthma were less pronounced. Two pathways identified in CD patients were also increased in asthma patients (P 0.001 in both sexes): [THAKAR_PBMC_INACTIVATED_INFLUENZA_AGE_21_30YO_RESPONDERS_28DY_DN] and [GSE3039_CD4_TCELL_VS_B1_BCELL_UP].

Conclusions: Our findings highlight a shared genetic risk between asthma and CD. Two shared gene sets related to altered PBMC activity and CD4+ T-cells were identified. These findings provide valuable insights into these overlapping mechanisms and identify targets for therapeutic interventions.

4.9. The Prevalence of Anti-Nuclear Autoantibodies in a Population Exposed to Natural Asbestiform Fibers: A Cross-Sectional Study

Serena Matera ¹, Davide Matera ², Lucia Rapisarda ³, Venerando Rapisarda ¹, Filetti Veronica ¹, Ermanno Vitale ⁴, Francesca Vella ¹

^1. 

Occupational Medicine, Department of Clinical and Experimental Medicine, University of Catania, 95124 Catania, Italy

^2. 

Mediterranean Oncology Institute, Viagrande, Italy

^3. 

Cannizzaro Hospital, Catania, 95123, Italy

^4. 

Departmente of Medicine and Surgery, University of Enna “Kore”, Piazza dell’Università, 94100 Enna EN

Fluoro-edenite (FE) is an asbestiform mineral fiber identified in lava rocks extracted from a quarry in Biancavilla, Italy. The material derived from these rocks has historically been utilized for construction purposes in the area, exposing the local population to potential health risks. Previous research has demonstrated that asbestos exposure can trigger autoimmune responses, including an increased frequency of anti-nuclear autoantibodies (ANAs). This study aimed to investigate the potential link between exposure to FE and autoimmune reactions in a population at risk.

A total of 120 participants, comprising 60 residents of Biancavilla and 60 individuals from a control group, were randomly selected for inclusion. All of the participants underwent complimentary medical evaluations, including spirometry and high-resolution computed tomography (HRCT) chest scans, to assess potential respiratory or pleural abnormalities. Their ANA levels were determined via indirect immunofluorescence, a reliable method for detecting autoimmune activity. No clinical signs or symptoms of illness were observed in any participant during the medical examination. ANA positivity was detected in 70% (n = 42) of the exposed subjects, compared to 25% (n = 15) of the control group (p0.05). Pleural plaques (PPs) were identified in 21 (30%) of the exposed individuals and in 2 (3%) of the controls, with all the PP-positive subjects also testing positive for ANAs.

These findings suggest a significant association between FE exposure and elevated ANA levels, mirroring observations in populations exposed to asbestos fibers. Furthermore, the co-occurrence of PP and ANA positivity highlights the need for further exploration into the potential relationship between environmental exposure to asbestiform fibers and autoimmune dysregulation. This study provides a foundational step for future research into the health effects of FE exposure and its broader implications for public health.

4.10. The Specific Combinations of Cytokines and Tumor-Derived Factors Determine the Particular Immunosuppressive Properties of In Vitro Generated, Murine, Myeloid-Derived Suppressor Cells

Mona Sami Awad ^1,2, Aleksandra Sen’kova ¹, Marina Zenkova ¹, Oleg Markov ¹

^1. 

Institute of Chemical Biology and Fundamental Medicine SB RAS, Novosibirsk, Russia

^2. 

Faculty of Natural Sciences, Novosibirsk State University, Novosibirsk, Russia

MDSCs are heterogeneous populations of immature myeloid cells with potent immunosuppressive capabilities that play a significant role in tumor immunology and autoimmune diseases. These cells have emerged as critical regulators of immune responses, impacting disease progression and treatment outcomes. MDSCs use multiple mechanisms to suppress immune cells, primarily T-cells, and can be divided into two subsets, polymorphonuclear (PMN-MDSCs) and monocytic (M-MDSCs) ones, which resemble mature neutrophils and monocytes, respectively.

To facilitate MDSC investigation and develop effective MDSC-mediated therapies, it is essential to establish reliable methods for their in vitro generation. In our study, we compared six protocols for the in vitro generation of functional mouse MDSCs from bone marrow progenitors. The protocols included granulocyte–macrophage-colony-stimulating factor (GM-CSF), alone or in combination with interleukin-6 (IL-6) or granulocyte-colony-stimulating factor (G-CSF), with or without a tumor-conditioned medium (TCM) derived from B16-F10 melanoma. The obtained MDSCs were characterized by the morphology, phenotype, and gene expression of key immunosuppressive factors, as well as the in vitro suppression of T-cell proliferation.

All the tested protocols yielded approximately 25% M-MDSCs and 50% PMN-MDSCs. The protocols utilizing IL-6-generated MDSCs with reduced maturation and differentiation markers upregulated the expression of Arg1 and Nos1 mRNA, increased levels of Arg-1 and TGF-β proteins, and enhanced ROS production compared to the other protocols. All the protocols under study generated MDSCs that efficiently inhibited T-cell proliferation in vitro, with some advantage for the GM-CSF and G-CSF+GM-CSF protocols. Interestingly, combining the protocols with B16-F10-TCM reduced the immunosuppressive properties of the generated MDSCs.

These results provide valuable insights into the optimal conditions for the in vitro generation of MDSCs with specific immunosuppressive properties. This knowledge will contribute to the development of more effective MDSC-mediated therapeutic approaches for various disorders, including tumor-induced immunosuppression, transplant complications, and autoimmune and inflammatory diseases.

This research was funded by the Russian Science Foundation (Grant/Award Number: 19-74-30011) and the Russian state-funded project for ICBFM SB RAS (Grant/Award Number: 125012300659-6).

5. Biomedicine in Cardiovascular Diseases

5.1. Hormone-Independent Sex Differences in Proinflammatory and Adhesion Mediators in a Cellular Model of Atherosclerosis

Riwka Palant ¹, Edith Oscherowa ², Weam Karmid ², Elena Kaschina ³, Jennifer Kirwan ⁴, Sarah Trajkovski ⁴, Mario Lorenz ⁵, Maria Luisa Barcena ⁶

^1. 

Charité Universitätsmedizin Berlin, corporate member of Freie Universität Berlin and Humboldt-Universität zu Berlin Klinik für Kardiologie, Angiologie und Intensivmedizin, CCM Deutsches Herzzentrum der Charité

^2. 

Charité—Universitätsmedizin Berlin, corporate member of Freie Universität Berlin and Humboldt-Universität zu Berlin, Department of Geriatrics and Medical Gerontology

^3. 

Charité—Universitätsmedizin Berlin, Institute of Pharmacology, Max Rubner Center for Cardiovascular Metabolic Renal Research (MRC), Berlin, Germany. DZHK (German Centre for Cardiovascular Research), partner site Berlin

^4. 

Berlin Institute of Health at Charité—Universitätsmedizin Berlin, Metabolomics Platform, Berlin, Germany

^5. 

Charité Universitätsmedizin Berlin, corporate member of Freie Universität Berlin and Humboldt-Universität zu Berlin Klinik für Kardiologie, Angiologie und Intensivmedizin, CCM Deutsches Herzzentrum der Charité. DZHK (German Centre for Cardiovascular Research)

^6. 

Department of Urology, Eberhard Karl University of Tuebingen, Tuebingen, Germany

Introduction: Women are under-represented in both clinical studies and drug development. However, significant sex differences exist in the occurrence and progression of diseases, which are predominantly attributed to hormonal influences. This also applies to cardiovascular diseases. Failure to consider the sex of cells in basic research can lead to erroneous results and false conclusions. Therefore, in this project, we aimed to investigate hormone-independent sex differences at the cellular level using a cell model of atherosclerosis.

Methods: Male and female human umbilical vein endothelial cells (HUVECs) from twin pairs were treated with either TNF-α (10 ng/mL) or with supernatants from proinflammatory-stimulated male THP-1 and female HL-60 monocyte-like cells. The expression of the focal adhesion proteins paxillin, vinculin, talin, and tensin and the cell adhesion molecules VCAM1 and ICAM-1 was assessed using quantitative real-time PCR or Western blot analysis. In addition, the levels of secreted cytokines and chemokines were measured in the HUVEC cell culture supernatants via a Cytokine Multiplex Assay.

Results: The female HUVECs exhibited increased levels of the secreted proinflammatory cytokines IL-17a and IL-22, the anti-inflammatory cytokine IL13, the pro-angiogenic cytokine TGF-α, and the chemoattractant protein Eotaxin in the supernatant after treatment with TNF-α or with supernatants from proinflammatory-stimulated monocyte-like cells compared to the controls (p < 0.05). Conversely, the male supernatants exhibited increased levels of IL17f and RANTES. Moreover, the cell adhesion proteins VCAM-1 and ICAM-1, the chemokine MDC, and the cytokine MIP-1α showed opposite directions of regulation between the female and male cells after both stimulations. The gene and protein expressions of the focal adhesion proteins paxillin, vinculin, talin, and tensin showed no significant differences.

Conclusions: These results provide evidence for the differential regulation of adhesion proteins and cytokines in female and male cells. Hormone-independent sex differences in endothelial cell signaling within a proinflammatory environment may be of relevance for the development of atherosclerosis.

5.2. Efficacy of Liposome-Encapsulated Doxycycline Against Methicillin-Resistant Staphylococcus aureus

Enkhtaivan Erdenee ^1,2, Baatarmanlai Dorjgochoo ², Odonchimeg Munkhjargal ³, Adilzaya Dashdorj ¹, Enkhjargal Dorjbal ¹, Baatarkhuu Oidov ¹, Ariunsanaa Byambaa ¹

^1. 

Mongolian National University of Medical Sciences, Ulaanbaatar 14200, Mongolia

^2. 

Department of Biomedicine, Etugen University, Ulaanbaatar 14200, Mongolia

^3. 

Mongolian Academy of Science, Institute of Chemistry and Chemical Technology, Ulaanbaatar14200, Mongolia

Introduction: Antibiotic-resistant bacterial infections pose a growing global health threat and are projected to become the leading cause of mortality by 2050. Staphylococcus aureus (S. aureus), a common antibiotic-resistant pathogen, colonizes approximately two billion people, with 53 million carrying methicillin-resistant S. aureus (MRSA). The increasing prevalence of MRSA presents treatment challenges, particularly in resource-limited settings. In Mongolia, antibiotic consumption reached 40.48 defined daily doses (DDDs) per 1000 inhabitants per day in 2020, underscoring the need for alternative therapies. This study aimed to encapsulate doxycycline in liposomes and evaluate its antibacterial efficacy against S. aureus and MRSA.

Methods: Phospholipids were extracted from egg yolk and analyzed by thin-layer chromatography (TLC). Liposomes were prepared using the thin-film hydration technique with freeze–thaw cycles. Atomic force microscopy (AFM) characterized the liposomes’ morphology and size, while high-performance liquid chromatography (HPLC) quantified their doxycycline content. The minimum inhibitory concentration (MIC) of liposome-encapsulated doxycycline was determined via broth microdilution assays against S. aureus and MRSA.

Results: The phospholipid extraction yielded 32%, with TLC confirming phosphatidylcholine presence. AFM revealed an average liposome height of 41.15 nm (±4.9 nm) and a diameter of 153.07 nm (±70.48 nm), with a polydispersity index of 0.212. HPLC showed a retention time of 5.843 min, confirming high doxycycline purity (981 ppm). The MIC values for free doxycycline against S. aureus and MRSA were 8 μg/mL and 32 μg/mL, respectively, whereas liposome-encapsulated doxycycline reduced the MICs to 4 μg/mL (S. aureus) and 16 μg/mL (MRSA).

Conclusions: Liposomes with an average diameter of 153 nm were successfully produced. Encapsulated doxycycline demonstrated enhanced antibacterial activity, reducing MIC values by 50% for both S. aureus and MRSA compared to free doxycycline. These findings highlight liposomal drug delivery as a promising approach for combating antibiotic-resistant infections.

5.3. In Silico Evaluation of Diketopiperazine (DPK) Derivatives as Potential Inhibitors for G-Protein-Coupled Receptors (GPCRs)

Sepideh Jafari, Bojarska Joanna

• Chemistry Department, Institute of Ecological and Inorganic Chemistry, Technical University of Lodz, 116 Zeromskiego St, Lodz 90-924, Poland

Background: G-protein-coupled receptors (GPCRs) are a diverse group of membrane proteins that mediate critical physiological processes by converting extracellular signals into intracellular responses. The β2-Adrenergic Receptor (β2-AR), a key GPCR, plays a pivotal role in smooth-muscle relaxation, bronchodilation, and cardiovascular function, making it a therapeutic target for conditions such hypertension or asthma. Diketopiperazines (DPKs), as the simplest cyclic peptides, have emerged as promising scaffolds for inhibiting protein interactions and modulating receptor activity, offering a novel, appealing therapeutic approach with potentially fewer side effects compared to small-molecule inhibitors.

Methods: In this study, five DPK derivatives were obtained from PubChem and evaluated for their binding affinity to a 3D structure of β2-AR (PDB ID = 2RH1) through molecular docking studies using Autodock 4.6 and MGLTools. Each compound’s binding energy and hydrogen bond formation were assessed to determine their interaction efficiency.

Results: Among the five compounds, tryptophan–proline diketopiperazine (compound 3) exhibited the highest binding affinity, with a binding energy of −5.89 kcal/mol, and formed two hydrogen bonds. The enhanced interaction is attributed to the aromatic nature of tryptophan, which promotes strong π-π stacking interactions, and the rigidity of proline, which allows for optimal fitting within the receptor’s binding pocket. The hydrophobic interactions further stabilized the complex.

Conclusions: This study highlights that diketopiperazine (DPK) derivatives, particularly tryptophan–proline diketopiperazine, are promising inhibitors of the β2-Adrenergic Receptor (β2-AR), a key G-protein-coupled receptor (GPCR). The compound’s aromaticity and rigidity enhance receptor interaction, providing insights into the design of peptide-based inhibitors for β2-AR and other GPCR-related diseases, with potential for improved specificity and fewer side effects.

5.4. Natural Inhibitors of the β2-Adrenergic Receptor: A Computational Study for Targeting GPCR-Mediated Diseases

Sepideh Jafari, Joanna Bojarska

• Faculty of Chemistry, Lodz University of Technology, Lodz, Poland

G-protein-coupled receptors (GPCRs) are integral membrane proteins that mediate critical physiological processes by translating extracellular signals into intracellular responses. Among them, the β2-Adrenergic Receptor (β2-AR) plays a significant role in smooth-muscle relaxation, bronchodilation, and cardiovascular regulation, making it a primary target for treating conditions such as asthma, hypertension, and chronic obstructive pulmonary disease (COPD) [1]. While synthetic drugs are widely used to target β2-AR, they often lead to adverse effects and drug resistance, necessitating the exploration of alternative therapeutic approaches [2].

This study utilized molecular docking methods with AutoDock 4.6 to investigate the interactions of natural compounds, including ephedrine, quercetin, catechin, and resveratrol, with β2-AR. The receptor’s 3D structure was retrieved from the Protein Data Bank (PDB ID: 2RH1) [3], and docking studies evaluated the binding energies, hydrogen bonding, and other stabilizing interactions between the ligands and the receptor’s active site. Ephedrine was observed to form hydrogen bonds with key residues, consistent with previous studies [4]. Quercetin exhibited strong binding interactions with the β2-AR’s active site, confirming its potential as a natural inhibitor [5].

These findings highlight the promise of natural compounds in modulating GPCR activity, particularly β2-AR, as safe and effective alternatives to synthetic drugs. The application of computational methods like molecular docking further emphasizes their utility in identifying bio-inspired strategies for drug discovery targeting GPCR-related diseases.

5.5. Risk Factors for Stroke and Dementia After Coronary Artery Bypass Grafting (5-Year Observation)

Irina Danilovna Syrova, Olga A Trubnikova, Stanislaw E Semenov

• Research Institute for Complex Issues of Cardiovascular Diseases, Kemerovo, Russian Federation

Objective: We wished to study the state of the brain, the frequency of carotid artery stenosis, and the duration of hypertension in patients with ischemic strokes and dementia over the long-term period following coronary artery bypass grafting.

Methods: A total of 75 people were included, and their average age before surgery was 56 [52; 60] years old. The operation was performed with the help of artificial blood circulation. The mean duration of the cardiopulmonary bypass was 86.0 [65.0; 105.0] min, and the surgery duration was 230.0 [190.0; 270.0] min. The follow-up period included the hospital stay and extended to the five-year stage. Neuroimaging was performed on a Somatom Sensation 64 multispiral computed tomograph (MSCT) (Siemens, Munich, Germany). Color duplex scanning of the brachycephalic arteries was performed using an ultrasound scanner of the expert class: “General Electric Vivid 7 Dimension LCD”, USA.

Results: Within five years after coronary artery bypass grafting, four (5.3%) of the patients developed strokes, and two (2.7%) of the patients developed dementia. The duration of arterial hypertension in the group before coronary artery bypass grafting was 4 [1; 10] years. In the patients with strokes, it was 8 [6; 15] years, and for those with dementia, it was 30 [20; 40] years. Before surgery, carotid artery stenoses of up to 50%, according to the criteria of NASCET (North American Symptomatic Carotid Endarterectomy Trial), were detected in 37% of them, while after five years, they were found in 55% of the patients, at p = 0.03. Among the stroke survivors, carotid artery stenosis was detected in four (100%) cases and dementia in one (50%) patient. Five years later, the expansion of the III ventricle of the brain was detected, in a range from 7.0 [5.5; 8.0] to 7.5 [6.0; 9.0] mm (p = 0.03). The number of people with leukoaraiosis increased from 25% to 71%, p = 0.0001, and those with cysts and gliosis increased from 4% to 20%, p = 0.0001, indicating damage to the small cerebral arteries.

Conclusions: It can be assumed that the presence of prolonged arterial hypertension and carotid artery stenoses increases the likelihood of strokes and dementia within five years after coronary bypass surgery.

5.6. Sepsis-Associated Aromatic Metabolites: Universal Markers of Bacterial Inflammation and Mortality in High-Risk Patients

Alisa Pautova, Natalia Beloborodova

• Federal Research and Clinical Center of Intensive Care Medicine and Rehabilitology, Moscow, 107031 Russia

Introduction: Postoperative infectious complications, especially in critically ill patients, have a high mortality risk. The early diagnosis of such complications and mortality prediction in the intensive care unit (ICU) are urgent tasks.

Objective: We aimed to identify and measure tyrosine and phenylalanine metabolites, which are clinically significant for the diagnosis of infectious complications, and to carry out mortality prediction in critically ill patients.

Materials and methods: A total of 298 patients were examined, including patients with acute abdominal surgical diseases (n = 58); patients who recently underwent cardiac surgery (n = 79) or neurosurgery (n = 82); and other high-risk patients on the day of admission to the ICU, regardless of the main pathology (n = 79). The level of aromatic metabolites in the blood serum or cerebrospinal fluid samples of the patients was determined by means of gas chromatography.

Results: In the patients with acute surgical diseases of the abdominal cavity with bacterial–inflammatory complications (n = 35), the median serum levels of phenyllactic (PhLA) and 4-hydroxyphenyllactic acid (p-HPhLA) were 2.5 (p < 0.001) and 1.5 (p = 0.048) times higher, respectively, than those of the patients without an infectious process (n = 23). Six hours after cardiac surgery, in the patients who subsequently developed infectious complications (n = 26), the median serum levels of p-HPhLA and the sum of sepsis-associated metabolites were 1.4 (p = 0.010) and 1.6 (p = 0.002) times higher, respectively, than those of the patients who did not develop complications (n = 36). In the post-neurosurgical patients with signs of secondary meningitis (n = 30), the median level of p-HPhLA in the cerebrospinal fluid was 2.7 times higher (p < 0.001) than that of the patients without signs of secondary meningitis (n = 52). On admission to the ICU, the patients who did not survive (n = 35) had at least 5-fold higher median serum levels of p-HPhLA and other sepsis-associated metabolites (p < 0.001) compared with those who survived (n = 44).

Conclusions: The levels of sepsis-associated metabolites of tyrosine and phenylalanine can be used to predict the development of infectious complications and mortality in high-risk patients.

5.7. Stabilized Curcumin in Emulsions and Hydrogel Systems: A Nutritional Strategy for Cardiovascular and Renal Protection in Hypercholesterolemia

Mihai Domnutiu Suciu ¹, Paul Andor ², Alexandra Trif ³, Mălina Suciu Petrescu ⁴, Monica Trif ², Alexandru Vasile Rusu ⁵

^1. 

Department of Urology, Clinical Institute of Urology and Kidney Transplant, “Iuliu Hatieganu” University of Medicine and Pharmacy, 400066 Cluj-Napoca, Romania

^2. 

University of Agricultural Sciences and Veterinary Medicine, 400372 Cluj-Napoca, Romania

^3. 

University of Oradea, 410087 Oradea, Romania

^4. 

Clinica Suciu Medical, 400331 Cluj-Napoca, Romania

^5. 

CENCIRA Agrofood Research and Innovation Centre, Ion Meșter 6, 400650 Cluj-Napoca, Romania

Hypercholesterolemia is a significant risk factor for cardiovascular diseases (CVDs) and has been increasingly associated with renal dysfunction. Nutritional interventions featuring bioactive compounds, such as curcumin, have gained attention due to their lipid-lowering, anti-inflammatory, and nephroprotective effects. A comprehensive literature search was conducted across the PubMed, Scopus, and Web of Science databases, selecting randomized controlled trials (RCTs), cohort studies, and systematic reviews published between 2010 and 2024. A meta-analysis of the selected studies demonstrated that curcumin supplementation significantly reduced total cholesterol (mean difference: −15.8 mg/dL, 95% CI: −20.4 to −11.2), triglycerides (−13.2 mg/dL, 95% CI: −18.5 to −7.9), and LDL levels (−10.5 mg/dL, 95% CI: −15.7 to −5.3), while increasing HDL levels (+6.7 mg/dL, 95% CI: 3.2 to 10.2). A systematic review and meta-analysis by Emami et al. [6] highlighted curcumin’s anti-inflammatory response in CKD patients. However, curcumin’s poor aqueous solubility and rapid metabolism limit its therapeutic potential. To overcome these challenges, researchers have developed advanced delivery systems, such as microencapsulation, emulsions, and hydrogel matrices, to enhance curcumin’s bioavailability, stability, and controlled release. Delivery systems, such as microencapsulation, nanoemulsions, and hydrogel matrices, enhanced curcumin’s bioavailability and therapeutic efficacy. Studies, including one carried out by Ashraf et al. [7], have highlighted the role of microencapsulated curcumin in modulating diet-induced hypercholesterolemia, supporting its potential as a therapeutic intervention.

Acknowledgments: Part of the work was supported by a grant from the Romanian Ministry of Education and Research, CCCDI-UEFISCDI, project number PN-IV-P1-PCE-2023-1092.

5.8. The Stimulation of the Auricular Vagus Nerve in ST-Elevation Myocardial Infarction (STEMI)

Sofia Kruchinova ^1,2, Vladimir Shvartz ³, Milana Gendugova ², Maria Karibova ², Alim Namitokov ^1,2, Maria Sokolskaya ³, Elena Kosmacheva ²

^1. 

Scientific Research Institute of Regional Clinical Hospital #1 Ochapovsky, Krasnodar, Russia

^2. 

Department of Therapy #1, Kuban State Medical University, Krasnodar, Russia

^3. 

Bakulev National Medical Research Center for Cardiovascular Surgery, Moscow, Russia

Over the past decade, it has been shown that transcutaneous vagus nerve stimulation (tVNS) has a cardioprotective effect, both in chronic heart failure and in coronary heart disease, preventing reperfusion injury and weakening myocardial remodeling.

The purpose of this study was to evaluate the prognostic effect of tVNS on hospital and long-term clinical outcomes in patients with STEMI.

Methods: A randomized, placebo-controlled trial. tVNS was performed on a group of patients who were eligible for the study according to the inclusion/exclusion criteria. tVNS was performed from the moment of admission to the PCI, during the PCI, and for the next 30 min after it. This clinical trial is registered with the ClinicalTrials database under a unique identifier: NCT05992259.

Results: A total of 109 patients were included: 54—Active tVNS; 55—Sham tVNS. At the hospital stage, the levels of myocardial damage markers (troponin, CPKMB, NTproBNP) in dynamics (after 6, 72, and 96 h) were significantly lower (p < 0.001) in the Active tVNS group. In this group, there were lower incidences of cardiac arrhythmias (p < 0.001) and cardiogenic shock (p = 0.044), and there was a better survival rate in the Active tVNS group compared to the Sham tVNS group (p = 0.024). After 12 months, in the long term, trends towards differences between the groups were found, without statistical significance. When comparing the patient groups for survival by Kaplan–Meier, we obtained statistically insignificant log-rank tests in terms of the total mortality (p = 0.618), in terms of the frequency of recurrent MI (p = 0.161), in terms of the frequency of hospitalization (p = 0.391), and in terms of the frequency of stroke (p = 0.490).

Conclusions: The use of tVNS in STEMI patients reduces myocardial damage, thereby reducing the incidence of hospital complications and hospital mortality. In the long term, there were trends towards differences in the groups in the frequency of MACE.

5.9. The Activator Protein-1 Complex Governs a Vascular Degenerative Transcriptional Program in Smooth Muscle Cells That Triggers Aortic Dissection and Rupture

Yongting Luo

• Department of Nutrition and Health, China Agricultural University, China

Background and aims: Stanford type A aortic dissection (AD) is a degenerative aortic remodeling disease marked by an exceedingly high mortality rate without effective pharmacologic therapies. Smooth muscle cells (SMCs) lining tunica media adopt a range of states, and their transformation from contractile to synthetic phenotypes fundamentally triggers AD. However, the underlying pathomechanisms governing this population shift and subsequent AD, particularly at distinct disease temporal stages, remain elusive.

Methods: Ascending aortas from nine patients undergoing ascending aorta replacement and five individuals undergoing heart transplantation were subjected to single-cell RNA sequencing. The pathogenic targets governing the phenotypic switch of SMCs were identified by trajectory inference, functional scoring, single-cell regulatory network inference and clustering, regulon, and interactome analyses and were confirmed using human ascending aortas, primary SMCs, and a β-aminopropionitrile monofumarate-induced AD model.

Results: The transcriptional profiles of 93,397 cells revealed a dynamic temporal-specific phenotypic transition and a marked elevation of the activator protein-1 (AP-1) complex, actively enabling synthetic SMC expansion. Mechanistically, tumor necrosis factor signaling enhanced AP-1 transcriptional activity by dampening mitochondrial oxidative phosphorylation (OXPHOS). Targeting this axis with the OXPHOS enhancer coenzyme Q10 or AP-1-specific inhibitor T-5224 impedes phenotypic transition and aortic degeneration while improving survival by 42.88% (58.3–83.3% for coenzyme Q10 treatment), 150.15% (33.3–83.3% for 2-week T-5224 treatment), and 175.38% (33.3–91.7% for 3-week T-5224 treatment) in the β-aminopropionitrile monofumarate-induced AD model.

Conclusions: This cross-sectional compendium of the cellular atlas of human ascending aortas during AD progression provides previously unappreciated insights into a transcriptional program permitting aortic degeneration, highlighting a translational proof of concept for an anti-remodeling intervention as an attractive strategy to manage temporal-specific AD by modulating the tumor necrosis factor-OXPHOS-AP-1 axis.

5.10. The Effect of the Short-Term Transcutaneous Electrical Stimulation of the Auricular Vagus Nerve on Parameters of Heart-Rate Variability

Vladimir Shvartz ¹, Eldar Sizhazhev ¹, Maria Sokolskaya ¹, Sofia Kruchinova ^2,3, Soslan Enginoev ⁴, Elena Shvartz ⁵

^1. 

Bakulev National Medical Research Center for Cardiovascular Surgery, Moscow, Russia

^2. 

Scientific Research Institute of Regional Clinical Hospital #1 Ochapovsky, Krasnodar, Russia

^3. 

Department of Therapy #1, Kuban State Medical University, Krasnodar, Russia

^4. 

Federal Center for Cardiovascular Surgery, Astrakhan, Russia

^5. 

National Medical Research Center for Therapy and Preventive Medicine, Moscow, Russia

Relevance: Previous studies have shown that transcutaneous vagus nerve stimulation (tVNS) can have a therapeutic effect similar to its invasive counterpart. However, an objective assessment of tVNS requires a reliable biomarker of successful vagus nerve activation.

The purpose of our study was to study the effect of the short-term, noninvasive stimulation of the auricular branch of the vagus nerve on heart-rate variability (HRV) parameters.

Methods: Patients were randomized into two groups according to the 1:1 scheme. Active tVNS was performed, attached to the tragus of the left ear. Sham tVNS was performed, attached to the earlobe of the left ear. The stimulation frequency was 20 Hz with a pulse duration of 200 microseconds. The research algorithm included four five-minute time intervals for recording biological signals: (1) initially at rest, (2) during the first 5 min of stimulation, (3) during the next 5 min of stimulation, and (4) after the end of stimulation. The HRV parameters evaluated in this study included the standard temporal and spectral characteristics of HRV. This clinical trial is registered with the ClinicalTrials database under a unique identifier: NCT05680337.

Results: A total of 111 patients were included in the study: 53 Active tVNS; 58 Sham tVNS. Initially, there were no differences in the HRV parameters between the groups. After the start of the stimulation, the parameters pNN50, NN50, RMSSD, VLF%, IC1, IC2, VLF%, HF%, and HF began to differ significantly between the groups. After the end of the stimulation, it was found that the RMSSD and HF% parameters were significantly lower in the Active tVNS group, and LF/HF were significantly higher in the Active tVNS group than in the Sham tVNS group.

Conclusions: After the start of the tVNS, there was a decrease in the levels of HRV parameters, reflecting the activity of the parasympathetic nervous system in the Active tVNS group. At the same time, there were no statistically significant dynamics in the Sham tVNS group.

5.11. The Protective Role of the SGLT2 Inhibitor Empagliflozin in an Endothelial Inflammation Model

Haiyan Wu ^1,2, Elisabeth Strässler ^3,4, Elena Kaschina ^4,5, Mario Lorenz ^1,4, Yury Ladilov ⁶, Maria Luisa Barcena ⁷

^1. 

Charité–Universitätsmedizin Berlin, corporate member of Freie Universität Berlin und Humboldt-Universität zu Berlin, Klinik für Kardiologie, Angiologie und Intensivmedizin CCM, Deutsches Herzzentrum der Charité

^2. 

Respiratory Department of Zhejiang Hospital, Hangzhou, China

^3. 

Charité–Universitätsmedizin Berlin, corporate member of Freie Universität Berlin und Humboldt-Universität zu Berlin, Klinik für Kardiologie, Angiologie & Intensivmedizin, Campus Benjamin Franklin, Deutsches Herzzentrum der Charité

^4. 

DZHK (German Centre for Cardiovascular Research), partner site Berlin, Germany

^5. 

Charité—Unversitätsmedizin Berlin, Institute of Pharmacology, Max Rubner Center for Cardiovascular Metabolic Renal Research (MRC), Berlin, Germany

^6. 

Department of Cardiovascular Surgery, Heart Center Brandenburg, Brandenburg Medical School, Bernau bei Berlin, Germany

^7. 

Department of Urology, Eberhard Karl University of Tuebingen, Tuebingen, Germany

Introduction: Chronic inflammation and mitochondrial dysfunction are key drivers of endothelial aging and cardiovascular diseases. Sodium–glucose co-transporter 2 (SGLT2) inhibitors are known for their cardiovascular protective effects. However, the underlying molecular mechanisms, especially their impact on autophagy and mitochondrial function, remain unclear. This study aims to investigate the effects of SGLT2 inhibitors on mitochondrial function and autophagy in endothelial cells.

Methods: Human endothelial cells derived from induced pluripotent stem cells (iPS-ECs) (n = 12) were activated with 10 ng/mL of TNF-α for 24 h to create a proinflammatory environment and were then treated with 1 µM Empagliflozin for an additional 24 h. Western blot analyses were performed to assess the expression and activity of key mitochondrial and autophagy markers, e.g., AMPK activity, SOD2 expression, and autophagy flux.

Results: The proinflammatory treatment with TNF-α resulted in mitochondrial dysfunction and a disruption of autophagy. AMPK, a key cellular energy sensor and regulator of mitochondrial homeostasis, showed no significant differences in the pAMPK/AMPK ratios among the investigated groups. SOD2, an anti-oxidative enzyme critical for mitochondrial antioxidant defense, exhibited lower expression in the TNF-α-treated cells compared to the untreated cells. Empagliflozin treatment partially recovered the expression of SOD2. Conversely, the autophagy flux (LC3II/I ratio) was elevated after the proinflammatory treatment but subsequently decreased following the Empagliflozin treatment.

Conclusions: Empagliflozin exhibits protective effects on endothelial inflammation by modulating mitochondrial function and autophagy activity. While Empagliflozin partially restored SOD2 expression, the AMPK activity was not changed. Additionally, SGLT2 inhibition recovered the autophagy flux to the basal levels, suggesting a regulatory effect of Empagliflozin on autophagy. These findings indicate that SGLT2 inhibitors may play a role in mitigating inflammation-induced mitochondrial dysfunction and autophagy disruption, offering potential therapeutic benefits for endothelial aging and cardiovascular diseases.

6. Diabetes, Obesity, and Metabolic Diseases

6.1. Biomedical Approaches Involving the Carob Tree (Ceratonia siliqua L.) for the Management of Obesity and Diabetes

Yassine Mouniane ¹, Mounia Mezouara ², Abdelaati Soufiani ², Meryem Doubi ², Ahmed Chriqui ², Issam El-Khadir ², Khadija Manhou ², Driss Hmouni ²

^1. 

Laboratory of Natural Resources and Sustainable Development, Faculty of Sciences, Ibn Tofaïl University—KENITRA-University Campus, Kenitra 14000, Morocco

^2. 

Natural Resources and Sustainable Development laboratory, Faculty of Sciences, Ibn Tofail University, B.P 242, Kenitra, Morocco

The carob tree (Ceratonia siliqua L.), a traditional Mediterranean plant, stands out as a promising natural solution in the fight against obesity and diabetes, two major public health issues worldwide. This plant is rich in dietary fiber, polyphenols, and various bioactive compounds that act synergistically to prevent and reduce the metabolic imbalances associated with these pathologies. Among its assets, carob promotes satiety, reduces fat absorption, and positively modulates intestinal microbiota, key mechanisms for improving weight management and blood-sugar regulation.

On the metabolic front, carob extracts help regulate blood sugar and insulin, thereby limiting the complications associated with obesity, such as type 2 diabetes and cardiovascular disease. The underlying biological mechanisms include the inhibition of the digestive enzymes responsible for the breakdown and absorption of carbohydrates and lipids, resulting in improved glycemic stability and a significant reduction in caloric intake. The dietary fibers contained in carob play an important satiety-enhancing role, reducing the sensation of hunger and promoting better food management.

In addition, the modulation of the intestinal microbiota by carob bioactive compounds improves carbohydrate and lipid metabolism, enhancing energy balance. Its powerful antioxidant properties reduce oxidative stress and chronic inflammation, two key factors in the pathophysiology of obesity and diabetes. Carob also acts on fat metabolism by stimulating lipolysis (the breakdown of stored fat) and inhibiting lipogenesis (the formation of new fat).

These multiple properties position the carob tree as an essential resource for the development of natural and sustainable therapies against obesity, diabetes, and their associated complications. This study highlights the potential biomedical applications of carob extracts, offering an innovative and ecological perspective for the prevention and management of these metabolic disorders in an integrative and sustainable approach.

6.2. Exploring the Role of Oral Antidiabetic Medications as Adjuncts in Depression Treatment

Marlene Santos ¹, Angela Santos ^2,3, Angelo Jesus ¹

^1. 

REQUIMTE/LAQV, Escola Superior de Saúde, Instituto Politécnico do Porto, Rua Dr. António Bernardino de Almeida, 4200-072, Porto, Portugal

^2. 

Escola Superior de Saúde, Instituto Politécnico do Porto, Rua Dr. António Bernardino de Almeida, 4200-072, Porto, Portugal

^3. 

Farmácia Moutinho, Largo da Boavista, 4860-501 Cabeceiras de Basto, Portugal

Depression is a complex and multifaceted disorder with a poorly defined etiology and numerous risk factors. Despite the availability of several antidepressant classes, many treatments have limitations that compromise their effectiveness and impact patients’ quality of life. Depression and diabetes share pathophysiological mechanisms, such as insulin resistance, inflammation, and neuroplasticity. This review aims to explore the potential of oral antidiabetic therapy as an alternative or adjunct treatment for depression by examining preclinical studies to understand its mechanisms of action and clinical studies to evaluate its therapeutic efficacy. A bibliographic search was conducted in the PubMed database, and articles were selected using defined inclusion and exclusion criteria. The selected studies were categorized into two groups: preclinical studies and clinical studies. A detailed analysis was conducted for each study using a structured reading form. Key findings highlighted Metformin and Pioglitazone as the most studied drugs with promising effects. Preclinical studies showed that Metformin reduced depression-like behaviors in animal models, and when combined with conventional antidepressants, it enhanced their therapeutic effect. Similarly, Pioglitazone demonstrated significant antidepressant properties by alleviating depressive symptoms in clinical settings, particularly in patients with concurrent depression and diabetes. The mechanisms identified in these studies, including the modulation of insulin resistance and improvements in neuroplasticity, provide insight into the efficacy of oral antidiabetic therapy for depression. These findings suggest a dual benefit for patients with comorbid diabetes and depression. While the evidence supports the potential role of these therapies, further research is required to optimize treatment protocols, investigate long-term outcomes, and expand the understanding of the underlying biological pathways.

6.3. A Systematic Review of the Potential Applications of Nutrigenomics to Prevent and Alleviate Metabolic Disorders

A. Perez-Vazquez ¹, P. Barciela ¹, A.O.S Jorge ^1,2, M. Carpena ¹, A.G. Pereira ^1,3, Ezgi Nur Yuksek ¹, M.A. Prieto ¹

^1. 

Universidade de Vigo, Nutrition and Bromatology Group, Department of Analytical Chemistry and Food Science, Instituto de Agroecoloxía e Alimentación (IAA)—CITEXVI, 36310 Vigo, Spain

^2. 

REQUIMTE/LAQV, Department of Chemical Sciences, Faculty of Pharmacy, University of Porto, R. Jorge Viterbo Ferreira 228, 4050-313 Porto, Portugal

^3. 

Investigaciones Agroalimentarias Research Group, Galicia Sur Health Research Institute (IIS Galicia Sur). SERGAS-UVIGO, Vigo, Spain

Metabolic disorders (MDs) are characterized by disruptions in normal metabolic processes, often leading to impaired carbohydrate, lipid, or protein metabolism. These disorders, including diabetes mellitus, phenylketonuria, and lysosomal storage diseases, occur due to genetic mutations, environmental influences, or their interplay. In recent decades, nutrigenomics (NG) has arisen as a promising field, in which nutrition plays a key role in understanding the interactions between nutrients and gene expressions. By elucidating the molecular mechanisms linking dietary components to metabolic pathways, NG enables the development of personalized nutrition strategies that could prevent or alleviate MDs. For instance, individuals with type 2 diabetes may benefit from dietary modifications targeting genes that are involved in insulin signaling pathways, while those with phenylketonuria require tailored diets that are low in phenylalanine. Advances in NG technologies, such as transcriptomics and epigenomics, have revealed how nutrients influence gene expression through different mechanisms, including DNA methylation and histone modification. These findings underscore the importance of dietary patterns in mitigating the risk of MDs and improving disease outcomes. Furthermore, NG contributes to precision medicine by identifying genetic polymorphisms that affect nutrient metabolism and enabling interventions that are aligned with an individual’s genetic profile. As research progresses, the integration of NG into clinical practice holds potential to revolutionize the prevention and management of metabolic disorders, promoting improved health outcomes. This systematic review aims to compile and analyze the current evidence on the role of NG in the prevention of MDs, focusing on its potential application in precision and preventive medicine. By synthesizing recent findings, this review highlights opportunities and limitations for integrating NG insights into clinical practice, helping lay the groundwork for innovative strategies to mitigate the global burden of metabolic disorders.

6.4. A Bioinformatic Analysis of Retatrutide’s Impact on Galectin-3 Signaling Networks in Type 2 Diabetes Mellitus: A Systems Biology Approach

Maul Shree ¹, Dhiraj Kishore ²

^1. 

Department of Medicine, Institute of medical sciences, Banaras Hindu University, Varanasi 221005

^2. 

Department of Gen. Medicine, IMS, Banaras Hindu University, Varanasi 221005

Introduction: Galectin-3, a β-galactoside-binding protein, plays a crucial role in inflammation and insulin resistance in type 2 diabetes mellitus (T2DM). This study investigated how Retatrutide, a novel GLP-1/GIP/glucagon receptor triple agonist, modulates the Galectin-3 pathway. Recent advances in multi-omics data analysis have highlighted the complexity of Galectin-3 signaling in type 2 diabetes mellitus (T2DM). This bioinformatic study employed network analysis and pathway modeling to elucidate how Retatrutide, a novel triple receptor agonist, influences the Galectin-3 regulatory network.

Methodology: We developed a computational framework integrating protein–protein interaction networks, pathway enrichment analysis, and dynamic modeling. Gene expression profiles were analyzed. This study incorporated regulatory network construction focusing on LGALS3 and associated genes, pathway topology analysis, and temporal expression pattern modeling. Key pathway components were mapped using the directed graph theory and centrality metrics.

Results: The network analysis identified five major regulatory hubs within the Galectin-3 pathway, with LGALS3 showing high betweenness centrality (0.85). Pathway enrichment revealed the significant modulation of inflammatory cascades (p < 0.001) and insulin signaling networks (p < 0.01). Temporal expression modeling demonstrated the biphasic regulation of LGALS3, with early suppression (−40%) followed by sustained downregulation. The graph theory analysis identified novel regulatory motifs linking GLP-1R activation to LGALS3 suppression through cAMP-dependent pathways.

Conclusions: Our bioinformatic approach revealed previously uncharacterized regulatory mechanisms in Retatrutide’s modulation of the Galectin-3 pathway. The network topology analysis suggests that Retatrutide’s effects are mediated through the coordinated regulation of multiple signaling nodes, rather than linear pathway suppression. These computational findings provide a systems-level understanding of Retatrutide’s mechanism of action and identify potential therapeutic targets for experimental validation.

6.5. In Silico Molecular Docking and ADMET Prediction of Ginkgo Biloba Biflavonoids as Dual Inhibitors of Human HMG-CoA Reductase and Alpha-Amylase

Nesteve John Agosto ^1,2

^1. 

Department of Chemistry, University of Science and Technology of Southern Philippines, Cagayan de Oro City 9000, Philippines

^2. 

Center for Natural Products Research, University of Science and Technology of Southern Philippines, Cagayan de Oro City 9000, Philippines

Phytochemicals are increasingly studied for their potential to inhibit HMG-CoA reductase (HMGR) and alpha-amylase enzymes, which are key targets in managing hypercholesterolemia and diabetes. The inhibition of HMGR reduces cholesterol synthesis, while alpha-amylase inhibition helps control postprandial blood glucose levels. This study used computational techniques, including molecular docking and ADMET (absorption, distribution, metabolism, Excretion, and toxicity) predictions, to evaluate the potential of five Ginkgo biloba biflavonoids—amentoflavone, bilobetin, ginkgetin, isoginkgetin, and sciadopitysin—as dual inhibitors of these enzymes. The molecular docking results demonstrated that amentoflavone (−10.1 kcal/mol) and bilobetin (−9.8 kcal/mol) exhibited stronger binding affinities to HMGR than the reference drug atorvastatin (−9.3 kcal/mol). Similarly, for α-amylase, amentoflavone (−11.5 kcal/mol), bilobetin (−11.3 kcal/mol), and ginkgetin (−11.1 kcal/mol) surpassed the binding affinity of the reference drug acarbose (−10.5 kcal/mol). These consistently strong binding affinities indicate that both amentoflavone and bilobetin have the potential to act as dual inhibitors of these two enzymes. Our ADMET analysis revealed that bilobetin demonstrated favorable drug-like properties, adhering to Lipinski’s rule of five, which predicts good oral bioavailability. Although bilobetin exhibited low gastrointestinal absorption, it was predicted to be non-mutagenic and non-hepatotoxic and demonstrated no significant toxicity risks, making it a highly promising candidate for further drug development. These important findings underscore the potential of Ginkgo biloba biflavonoids for addressing metabolic disorders. Future in vitro and in vivo studies are essential to validate these in silico results, providing deeper insights into their therapeutic applications and contributing to the development of novel dual-action drugs targeting hypercholesterolemia and diabetes.

6.6. Investigating the Anti-Obesity Effects of Kalanchoe pinnata

Maryam Ashraf, Kanwal Ashiq, Muhammad Zaeem Ahsan, Rimsha Ahsan

• Faculty of Pharmaceutical Sciences Superior College, Superior University Lahore, Pakistan

Introduction: Kalanchoe pinnata has long been recognized for its medicinal properties, especially in the treatment of metabolic diseases, like obesity. This study examines the phytochemical makeup of K. pinnata root–stem methanol extract and looks at its potential as a treatment for weight management.

Method: Using a thorough phytochemical analysis that combined qualitative and quantitative techniques, the bioactive components of the root–stem powder were identified and measured. Functional groups were determined using FTIR and UV–Vis spectroscopies. A GC-MS analysis was used to profile volatile chemicals. An in vitro pancreatic lipase inhibition experiment was used to evaluate the methanol extract’s anti-obesity efficacy. To find out how the extract affected lipid profiles and histopathological alterations in the pancreas and other organs, an in vivo investigation employing a high-fat diet was also carried out.

Results: The findings of the phytochemical screening revealed the presence of compounds such as flavonoids, polyphenols, saponins, and glycosaponins, which are known to enhance metabolic health. At the same time, the GC/Mass analysis detected essential compounds, such as different fatty acids and phytol octadecanoic acid. The K. pinnata extract in methanol markedly inhibited pancreatic lipase activity. After the extract treatment and a high-fat diet, the rats’ lipid profiles improved and displayed decreased LDL and total cholesterol levels. The restoration of the normal pancreatic liver and kidney architecture and the reduction of inflammation were found by histopathological examinations, and these effects were comparable to those of the orlistat.

Conclusions: The study’s findings demonstrate that K. pinnata has two key anti-obesity benefits: enhanced metabolic parameters and preventive effects on organ health. The results necessitate more research into the precise mechanisms of action and possible therapeutic uses of this plant extract in the treatment of obesity.

6.7. Maternal Metabolic Syndrome Impairs Offspring Neurodevelopment, Behavior, and Metabolism with Sexual Dimorphism

Ana Sofia Vallés, Facundo Prado Spalm, Marié Lucía Cuervo Sanchez, Natalia Furland

• Instituto de Investigaciones Bioquímicas de Bahía Blanca (INIBIBB)-CONICET, Universidad Nacional del Sur (UNS), Bahía Blanca 8000, Argentina

Introduction: Maternal metabolic syndrome (MetS) can profoundly impact offspring development. This study aimed to evaluate the effects of maternal MetS on somatic growth, developmental milestones, neurochemical and metabolic alterations, and behavior using an animal model.

Methods: Female Wistar rats were given 20% fructose in drinking water before and during pregnancy to induce MetS, while the control group consumed only water. Their offspring were assessed for biochemical markers in whole-brain tissue at postnatal day (PN) 1 and for somatic development and developmental milestones from birth to PN21. Behavioral tests evaluating anxiety-like behavior, cognition, nociception, and social interactions were conducted in adolescence and early adulthood. Their metabolic status was assessed at PN100.

Results: At PN1, the female MetS offspring exhibited increased lipid peroxidation (TBARS levels), heightened antioxidant enzyme activity (catalase) and expression (superoxide dismutase), and elevated proinflammatory COX-2 expression in whole-brain tissue. Neurodevelopmental delays were observed in both the females (delayed grasp reflexes, cliff aversion, plantar reflex, and righting reflex) and the males (delayed forelimb grasp and plantar reflex). Behaviorally, the female MetS offspring displayed increased anxiety-like behavior, impaired social interactions, heightened neophobia, and altered nociception, while the males exhibited cognitive deficits and social interaction impairments. Additionally, the female MetS offspring showed prolonged latency in the nocifensive tail flick response to thermal stimuli, indicating altered pain sensitivity. At PN100, the female MetS offspring exhibited increased visceral fat accumulation and elevated triglyceride levels, suggesting long-term metabolic disturbances.

Conclusions: Maternal MetS disrupts offspring development, leading to early biochemical imbalances, neurodevelopmental delays, and long-term metabolic dysfunction. These effects were sex-dependent, with females showing greater oxidative stress, inflammation, anxiety, and metabolic alterations, while males exhibited milder but significant neurodevelopmental and cognitive impairments. These findings underscore the long-lasting impact of maternal MetS on offspring health and highlight the need for early interventions to mitigate neurodevelopmental and metabolic risks.

6.8. Molecular Insights into Oxidative Stress in T2DM-Driven CAD: Clinical Implications and Biomarker Innovation

Shubhra Vats, Dhiraj Kishore

• Department of General Medicine, Institute of Medical Sciences, Banaras Hindu University, Varanasi, Uttar Pradesh 221005, India

Introduction: Type 2 diabetes mellitus (T2DM) affects 462 million people globally and is linked to complications like retinopathy, nephropathy, neuropathy, and coronary artery disease (CAD). T2DM increases the CAD risk, contributing to 75% of related mortality. Genetic predispositions and distinct mechanisms differentiate DM-CAD from other forms, such as CAD, caused by hypertension or degenerative changes, requiring gene expression profiling and transcriptome and reactome analyses to identify and establish molecular markers and improve its diagnosis, prognosis, and treatment strategies. Methods: The Gene Expression Omnibus (GEO) datasets GSE250283 and GSE90074 were used to identify the differentially expressed genes involved in DM-CAD. Biological targets were identified that coincided with the identified differentially expressed genes. The biological pathways were analyzed using KEGG, and hub genes were identified for a further functional and signaling pathway analysis.

Results: The differentially expressed gene biomarkers were identified between the control and DM and DM-CAD. NLRP3, TLR4, STAT3, IL6, TNF-α, and NF-κB were found to be upregulated, while PPARG, SIRT1, and ADIPOR1 were downregulated, indicating significant pathways with involvement, including the oxidative stress response, JAK-STAT signaling, and insulin resistance and mitochondrial dysfunction.

Conclusions: Oxidative stress emerges as a critical driver of T2DM-CAD pathogenesis, influencing inflammatory pathways and metabolic dysfunction. Genes such as SOD2, CAT, and GPX1 highlight disruptions in antioxidant defense mechanisms, aligning with mitochondrial dysfunction. Elevated STAT3 expression in the JAK-STAT pathway and NLRP3 activation further exacerbate oxidative damage and inflammation. Meanwhile, the downregulation of SIRT1 and ADIPOQ underscores impaired glucose regulation and insulin sensitivity. These findings position oxidative stress as a key therapeutic target, alongside inflammasome and immune signaling pathways, like the JAK-STAT pathway, as novel therapeutic targets for mitigating T2DM-CAD’s severity.

6.9. The Polyphenol-Mediated Activation of PPARγ and the Adiponectin Pathway for the Treatment of Type 2 Diabetes

Muhammad Safdar ¹, Yasmeen Junejo ²

^1. 

Department of Breeding & Genetics, Cholistan University of Veterinary and Animal Sciences Bahawalpur 63100, Pakistan

^2. 

Department of Physiology & Biochemistry, Cholistan University of Veterinary and Animal Sciences Bahawalpur 63100, Pakistan

Peroxisome proliferator-activated receptor gamma (PPARγ) is a crucial nuclear receptor in humans, playing a central role in regulating glucose homeostasis and lipid metabolism. Its activation holds significant potential not only for managing type 2 diabetes but also for preventing its onset by addressing insulin resistance, a key precursor to the disease. This study explores the potential of polyphenols as natural modulators of PPARγ, aiming to enhance strategies for preventing the progression of insulin resistance to type 2 diabetes. Polyphenols, plant-derived bioactive compounds, possess strong bioactivity in modulating various cellular pathways, including those linked to PPARγ activation. Since insulin resistance can develop years before the onset of type 2 diabetes, identifying compounds that enhance insulin sensitivity early is critical for disease prevention. Molecular docking studies were conducted on a library of 12,000 polyphenols to identify compounds with the strongest binding affinities for PPARγ. Among the candidates, Resveratrol and Epigallocatechin gallate (EGCG) emerged as top ligands, with docking scores of −9.38 kcal/mol and −8.77 kcal/mol, respectively, along with lower RMSD values (0.55 and 0.72). These compounds demonstrated strong binding to key receptor sites of PPARγ, potentially enhancing its activity and improving insulin sensitivity, thereby reducing the risk of type 2 diabetes development. Moreover, the activation of the adiponectin signaling pathway was identified as a crucial mechanism underlying the beneficial effects of PPARγ in mitigating insulin resistance. Molecular dynamics simulations (MDSs) further validated the stability and efficacy of polyphenol–PPARγ complexes, confirming their sustained interactions and enhanced receptor activation. The pharmacokinetic profiles of these polyphenols revealed highly favorable characteristics, such as excellent bioavailability and non-toxicity, reinforcing their potential as preventive agents. Collectively, this study provides new insights into the modulation of PPARγ by polyphenols, offering a promising approach for delaying or preventing type 2 diabetes through natural compounds that target early metabolic dysfunction.

6.10. The Role of Interferon Therapy in Stabilizing Metabolic Liver Diseases

Megala T ¹, Nandha Gopal T ¹, Siva Pradeep M ¹, Sivabalan M ¹, Yasotha A ², Raja N ^1,3

^1. 

Sri Ramakrishna Mission Vidyalaya College of Arts and Science, Coimbatore, Tamilnadu, India

^2. 

United Institute of Technology, Coimbatore, Tamilnadu, India

^3. 

Rathinam Technical Campus, Coimbatore, Tamil Nadu, India

Metabolic diseases, including diabetes, obesity, and insulin resistance, have emerged as significant global health challenges with profound implications for liver health. As a central organ in glucose and lipid metabolism, the liver is particularly vulnerable to disruptions caused by these diseases. Such disturbances often result in complications like non-alcoholic fatty liver disease (NAFLD) and non-alcoholic steatohepatitis (NASH), which are major contributors to liver-related morbidity. This study develops a mathematical model to explore the progression of these conditions and evaluate the role of interferon therapy in controlling disease dynamics.

The analysis focuses on two key equilibrium states: the disease-free equilibrium (DFE), which represents the absence of disease, and the endemic equilibrium (EE), where the disease persists in the population. A critical component of the study is the basic reproduction number (R0), which serves as a threshold indicator of whether the disease will spread or die out. By comparing scenarios with and without interferon therapy, the study demonstrates how the therapy significantly reduces the R0, shifting the system from an endemic state to a disease-free state under specific conditions.

The findings highlight the potential of interferon therapy to stabilize liver health and reduce the prevalence of liver-related complications associated with metabolic diseases. This research provides valuable insights into the conditions under which interferon therapy is most effective, offering practical guidance for optimizing treatment strategies and public health interventions. By addressing the underlying dynamics of these diseases, this study contributes to a deeper understanding of their progression and supports global efforts to alleviate their impact.

6.11. Variation in White Blood Cell Counts Among Individuals with Type 2 Diabetes Mellitus

Saheed Shiffana ¹, Hazeemdeen Rukaiya ²

^1. 

Lecturer, Department of Medical Laboratory Technology, School of Medical Laboratory Technology, Peradeniya 20400, Ministry of Health, Sri Lanka

^2. 

Student, Department of Biomedical Sciences, British College of Applied Studies (BCAS Campus), Colombo 03, 00300, Sri Lanka

Type 2 diabetes mellitus (T2DM) is a chronic metabolic disorder characterized by insulin resistance and hyperglycemia. Emerging evidence suggests that variations in white blood cell (WBC) counts among individuals with T2DM may provide insights into the inflammatory and immune responses associated with the disease. This study aims to investigate WBC count variations in T2DM and their relationship with glycemic control and other clinical parameters.

A cross-sectional study was conducted at Colombo Medi Lab, Bambalapitiya, Sri Lanka, involving 321 participants categorized into pre-diabetic (FBS: 100–125 mg/dL, postprandial blood glucose: 140–199 mg/dL; HbA1c: 5.7–6.4%) and diabetes (FBS > 126 mg/dL; postprandial blood glucose: >200 mg/dL; HbA1c: ≥6.5%) groups. WBC counts and demographic data were collected, and statistical analyses were performed using Microsoft Excel and SPSS 20.0, with a significance threshold of p < 0.05.

Among the 321 participants, 171 were T2DM patients (30 pre-DM, 113 females, 58 males), while 150 were healthy individuals (95 females, 55 males). Significant differences (p < 0.05) were observed between the DM, pre-DM, and control groups in the WBC count, neutrophil count, lymphocyte count, N/L ratio, MID count, and FBS.

The WBC, lymphocyte, and MID counts showed a significant (p < 0.001) positive correlation with age, whereas the neutrophil count, N/L ratio, and FBS showed a negative correlation. The N/L ratio and MID count positively correlated with gender, while the WBC count, lymphocyte count, and FBS were negatively correlated. The WBC count, neutrophil count, N/L ratio, and FBS correlated positively with the BMI, whereas the lymphocyte and MID counts were negatively correlated.

This study highlights WBC count variations as potential biomarkers for monitoring T2DM progression. Future research should focus on longitudinal and interventional studies to explore causal relationships and clinical applications in diabetes management.

7. Gene Therapy and Gene Editing

7.1. Reshaping the Tumor Immune Microenvironment Through ROS-Responsive Nanoparticles Featuring the CRISPRi System to Mediate Programmed Death-Ligand 1 Attenuation and Enhance Cancer Immunotherapy via an Epigenetic Inhibitor

Qianru Li ¹, Huan Deng ², Songwei Tan ¹

^1. 

Huazhong University of Science and Technology, Wuhan 430070, China

^2. 

Department of Chemistry, Chemical and Life sciences, Wuhan University of Technology, Wuhan 430070, China

The involvement of programmed death-ligand 1 (PD-L1) in tumor cells, with its receptor PD-1 on immune cells, can transmit an inhibitory signal to induce immune evasion. Although the immune checkpoint inhibitor PD-L1 antibody has achieved significant antitumor efficiency in clinical treatment, its wide clinical application still involves several side effects and individual selectivity. In our ongoing research, we intend to use the CRISPR interference (CRISPRi) system to suppress PD-L1 expression in tumor cells; in this study, we combined it with the epigenetic inhibitor azacytidine (AZA) for enhanced cancer immunotherapy. We first fabricated a reactive oxygen species (ROS)-responsive poly(β-amino ester) (PBAE)-S-AZA cationic polymer prodrug. The CRISPRi plasmid and the PBAE-S-AZA prodrug were complexed to form the final nanoparticles via electrostatic interaction. The nanoparticles were uptaken in tumor cells with high efficiency, followed by the escape of the plasmid via the endosome/lysosome with the cooperation of PBAE. The release of the CRISPRi plasmid can lead to the suppression of PD-L1 in tumor cells and the relief of the immune checkpoint blockade. In the meantime, the epigenetic inhibitor AZA was also released from the nanoparticles due to the high intracellular ROS level to synergistically enhance the efficacy of immunotherapy. It was demonstrated that the nanoparticles can significantly increase the proportions of CD8+ T-cells, CD4+ T-cells, and M1 macrophages in the tumor microenvironment while decreasing the proportion of regulatory T-cells (Tregs) and M2 macrophages. Thus, the nanoparticles help to achieve the combination of genome editing, immunotherapy, and epigenetic regulation effectively. This study provides a novel platform for promoting antitumor treatment and precision medicine.

7.2. Reshaping the Tumor Immune Microenvironment: ROS-Responsive Nanoparticles Delivering the CRISPRi System for Programmed Death-Ligand 1 Suppression Are Combined with an Epigenetic Inhibitor to Enhance Cancer Immunotherapy

Huan Deng ¹, Songwei Tan ²

^1. 

Department of Chemistry, Chemical and Life sciences, Wuhan University of Technology, Wuhan 430070, China

^2. 

Huazhong University of Science and Technology, Wuhan 430070, China

The engagement of programmed death-ligand 1 (PD-L1) on tumor cells with its receptor PD-1 on immune cells can transmit an inhibitory signal to induce immune evasion. Although the immune checkpoint inhibitor PD-L1 antibody has significant antitumor efficiency in clinical treatment, its widespread clinical application still faces several side effects and individual selectivity. In our research, we use the CRISPR interference (CRISPRi) system to suppress PD-L1 expression in tumor cells and combine it with the epigenetic inhibitor azacytidine (AZA) for enhanced cancer immunotherapy. We first fabricate a reactive oxygen species (ROS)-responsive poly(β-amino ester) (PBAE)-S-AZA cationic polymer prodrug. The CRISPRi plasmid and the PBAE-S-AZA prodrug are complexed to form the final nanoparticles via electrostatic interaction. The nanoparticles exhibit high efficiency in uptake by tumor cells, followed by the endosome/lysosome escape of the plasmid with the cooperation of PBAE. The release of the CRISPRi plasmid can lead to the suppression of PD-L1 in tumor cells and the relief of the immune checkpoint blockade. Meanwhile, the epigenetic inhibitor, AZA, is also released from the nanoparticles due to the high intracellular ROS level to enhance the efficacy of the immunotherapy synergically. It was demonstrated that the ROS-responsive nanoparticles could significantly increase the proportion of CD8+ T-cells, CD4+ T-cells, and M1 macrophages in a tumor microenvironment, while decreasing the proportions of regulatory T-cells (Treg) and M2 macrophages. Thus, the nanoparticles help to realize the combination of genome editing, immunotherapy, and epigenetic regulation effectively. Our method provides a novel platform for promoting antitumor treatment and precision medicine.

7.3. The Application of PEG Oligomer-Modified Poly(β-Amino Esters) in Enhancing Plasmid Transfection

Guan Yang ¹, Lai chongwu ²

^1. 

Zhejiang Chinese Medical University, Zhejiang Chinese Medical University, Hangzhou 310053, China

^2. 

Huazhong University of Science and Technology, Wuhan, China

Introduction: Gene therapy offers a promising approach for treating various diseases, yet its success hinges on efficient and safe gene delivery systems. Poly(β-amino esters) (PBAE) is an excellent non-viral gene carrier that has the advantages of easy synthesis and various chemical structures. However, there are some obvious shortcomings, such as its cytotoxicity and limited penetration in tissues. To address these issues, we developed PEGylated PBAE (PBAE-E) with tailored chain lengths and contents to enhance both its transfection and biocompatibility.

Methods: We synthesized PBAE-E by incorporating short-chain PEG into the PBAE structure via Michael addition polymerization. The materials were characterized using 1H-NMR and FT-IR. PBAE-E nanoparticles were formed by complexing with the GFP plasmid and were analyzed for size, morphology (DLS, TEM), and DNA encapsulation (agarose gel electrophoresis). The in vitro transfection was assessed in multiple cancer cell lines, and the cytotoxicity was evaluated using the MTT assay. The in vivo imaging of mouse vaginas was used to investigate whether the introduction of PEG chains could increase the mucosal permeability of nanoparticles.

Results: PBAE-E was synthesized successfully, as confirmed by 1H-NMR and FT-IR. The PBAE-E could compact the plasmid into spherical nanoparticles with a diameter of about 250 nm. The PBAE-E nanoparticles demonstrated improved transfection efficiency and reduced cytotoxicity compared to the unmodified PBAE. Notably, PBAE-E4-5 and PBAE-E9-5 exhibited the best performance. In vivo imaging in mice revealed the enhanced mucosal permeability of PBAE-E, suggesting improved tissue penetration.

Conclusions: Our study demonstrates that PEG-modified PBAEs can significantly enhance plasmid transfection efficiency and biocompatibility compared to unmodified PBAEs. The PEGylation strategy provides a promising avenue for improving non-viral gene therapy.

8. Rare Diseases

8.1. Therapeutic Potential of Natural Plant Derivatives Against Retinoblastoma

Manshi Rana ¹, Rupali Sharma ¹, Shekhar Sharma ²

^1. 

Amity University Haryana, Gurugram (Manesar), India

^2. 

Lloyd Institute of Management and Technology, Department of Pharmaceutics, Knowledge Park II, Greater Noida 20136, U.P, India

Retinoblastoma (RB) is a rare pediatric retinal cancer. Due to financial and medical limitations, many children with retinablastoma (RB) only receive therapy when the disease has progressed to the metaphase and advanced clinical stages, which increases the risk of blindness and impairment. Despite the fact that there are numerous methods for treating RB, some children do not respond well to treatment for a variety of reasons. Natural medicines produced from plants are frequently employed in research into RB and have demonstrated clear therapeutic effects in the treatment of a variety of malignancies. In order to offer suggestions for the clinical usage of these medications, as well as the creation of novel therapeutic pharmaceuticals, we examine natural compounds produced from plants that are employed in the investigation of anti-RB.

It has also been demonstrated that certain naturally occurring chemicals, in addition to conventional anticancer medications, are useful in treating RB. These consist of sterol derivatives (such as ursolic and oleanolic acid), naphthoquinones (such as β-lapachone), and catechol derivatives (such as curcumin). Our main objective is to explore natural novel therapeutics for RB and make use of these natural active moieties in the development of a nano drug delivery system.

8.2. The Identification of Key Long Non-Coding RNAs (LncRNAs) Affecting Alternative Splicing and Gene Expression in the Frontal Cortex of Myotonic Dystrophy Type 1 Patients

Tehreem Fatima

• University of Connecticut, Storrs, CT 06269, USA

Myotonic dystrophy type 1 (DM1) is a rare and complex genetic disorder characterized by the expansion of CTG repeats within the 3’ untranslated region of the dystrophia myotonic protein kinase (DMPK) gene. With a prevalence of approximately 10 per 100,000 people worldwide, DM1 presents with severe neuromuscular symptoms, including early-onset ataxia, dysarthria, muscle weakness, and exercise intolerance. The multisystemic nature of DM1 underscores the importance of exploring its molecular underpinnings to develop targeted therapeutic strategies.

In this study, I investigated the role of long non-coding RNAs (lncRNAs) in DM1 pathophysiology. Once dismissed as “genomic junk”, lncRNAs are now recognized as pivotal regulators of gene expression and alternative splicing, making them key candidates for understanding the mechanisms underlying DM1. My research identified NUTM2A-AS1 as a critical lncRNA influencing splicing patterns in DM1-affected brains. Notably, NUTM2A-AS1 exhibited non-coding repeat expansions, implicating it in DM1’s development and progression. Other lncRNAs were also found to be prevalent in splicing mechanisms in DM1.

Further, I identified additional lncRNAs, such as those associated with KHDRBS3 and HDAC2, that significantly impact alternative splicing. These findings provide unique insights into the regulatory networks involved in DM1. My analysis of differential gene expression highlighted several lncRNAs and their corresponding genes or proteins, including MAP6, FOSL2, and HLA-DQB1, all of which contribute to the disorder’s multifaceted pathology.

These discoveries advance our understanding of the molecular mechanisms underlying DM1, emphasizing the intricate interplay of lncRNAs, genes, and proteins. This research lays the groundwork for future studies aimed at developing targeted therapies to address the diverse manifestations of this debilitating disorder.

8.3. Navigating Life with Rare Syndromes and Congenital Heart Disease: The Unseen Link

Shadab Ahamad ¹, Prachi Kukshal ¹, Priti Sharma ¹, Paramvir Singh ²

^1. 

Sri Sathya Sai Sanjeevani Research Centre, Palwal, Haryana, India

^2. 

Sri Sathya Sai Sanjeevani International Centre for Child Heart Care & Research, Palwal, Haryana, India

Introduction: Congenital heart disease (CHD) is the leading birth anomaly and a major contributor to infant mortality, especially in low- and middle-income countries, including India. Its etiology is multifactorial, involving both genetic and environmental factors. Notably, approximately 30% of CHD cases are associated with genetic syndromes, which often present with extracardiac anomalies. This study seeks to explore rare genetic syndromes linked to CHD, with a focus on the socio-demographic, socio-economic, and clinical profiles of the affected families.

Method: A hospital-based population screening study for CHD, including clinical dysmorphism examinations, was conducted from 2018 to 2024 at the Sri Sathya Sai Sanjeevani International Center for Child Heart Care & Research, Haryana (India), a totally free-of-cost pediatric cardiac care center. Comprehensive data were collected at the Sri Sathya Sai Sanjeevani Research Centre and analyzed using SPSS software.

Results: A total of 442 syndromic cases were identified, with Down’s syndrome being the most common (61.7%). Rare syndromes included Noonan (17), Marfan (11), Ellis–Van Creveld (8), DiGeorge (6), Williams (6), Pentalogy of Cantrell (3), Treacher Collins (3), MRKH (2), TAR (2), Congenital facial Nerve Palsy (2), Goldenhar (2), Alagille (1), Cornelia de Lange (1), Heterotaxy (1), Holt Oram (1), etc. The most prevalent CHD phenotype observed was ventricular septal defects (24%). Geographically, a significant proportion of syndromic cases came from the highly populated Indian state of Uttar Pradesh (55.6%), with a large number of affected families from the upper lower-socio-economic class (Class IV; 46.6%).

Discussion: Identifying the clinical variability in syndromes associated with CHD can facilitate an early diagnosis, which is crucial for timely intervention and improved outcomes. This study also highlights the socio-economic disparities in access to care, emphasizing the need for increased healthcare resources in underserved regions. Understanding the genetic basis of phenotypic features may help reduce disease-related mortality and morbidity.

8.4. Neurobrucellosis Presenting as Pyrexia of Unknown Origin and Meningoencephalitis: A Rare and Challenging Case

Avani Mendpara ¹, Brijesh Koyani ²

^1. 

Consultant Physician at H J Doshi Hospital, Rajkot, Gujarat (India)

^2. 

Consultant Chest Physician at H J Doshi Hospital, Rajkot, Gujarat (India)

Background: Neurobrucellosis is a rare and potentially life-threatening complication of brucellosis, a zoonotic infection caused by Brucella species. It can present with a wide range of neurological symptoms, making its diagnosis challenging.

Case Presentation: We report a rare and challenging case of neurobrucellosis presenting as pyrexia of unknown origin (PUO) and meningoencephalitis in a 45-year-old male. The patient presented with a 2-month history of fever, headache, and confusion. Extensive investigations, including blood cultures, imaging studies, and serological tests, were inconclusive. Brucella serology was eventually performed, revealing high titers of anti-Brucella antibodies.

Diagnosis and Treatment: The patient was diagnosed with neurobrucellosis and treated with a combination of antibiotics, including doxycycline, rifampicin, and streptomycin. He showed significant clinical improvement, with the resolution of fever, headache, and confusion.

Discussion: This case highlights the importance of considering neurobrucellosis in the differential diagnosis of PUO and meningoencephalitis, particularly in patients with a history of exposure to animals or the consumption of unpasteurized dairy products. Early diagnosis and treatment are crucial to prevent long-term neurological sequelae and improve outcomes. A thorough medical history, physical examination, and diagnostic workup are essential for the early detection and treatment of this potentially life-threatening condition.

Conclusions: Neurobrucellosis is a rare and challenging diagnosis that requires a high index of suspicion. This case report emphasizes the importance of considering neurobrucellosis in the differential diagnosis of PUO and meningoencephalitis, and it highlights the need for prompt treatment to improve outcomes. Clinicians should be aware of the clinical manifestations and diagnostic challenges of neurobrucellosis to ensure its timely and effective management.

8.5. Pharmacological Approaches in Amyotrophic Lateral Sclerosis: A Systematic Review of Clinical Trials on Efficacy, Survival, and Safety Profiles

Pedro Tomé ¹, Fernando Moreira ², Marlene Santos ²

^1. 

Farmácia Avenida, Porto, Portugal

^2. 

REQUIMTE/LAQV, Escola Superior de Saúde, Instituto Politécnico do Porto, Rua Dr. António Bernardino de Almeida, 4200-072, Porto, Portugal

Amyotrophic lateral sclerosis is a progressive neurodegenerative disease that results in the deterioration of neuromuscular connections, with no effective treatment currently available. The present study aimed to synthesize clinical trial data regarding pharmacological approaches in amyotrophic lateral sclerosis, focusing on the impact of various drugs on the Amyotrophic Lateral Sclerosis Functional Rating Scale (ALSFRS-R), patient survival, and safety profiles. A systematic review was conducted following the PRISMA 2020 guidelines (PROSPERO ID: CRD42023373675). The inclusion criteria consisted of clinical trials published in Portuguese, Spanish, or English, with participants diagnosed solely with amyotrophic lateral sclerosis and no other comorbidities. The exclusion criteria were systematic reviews, meta-analyses, and studies involving patients with additional health conditions. A total of 616 articles were identified through databases such as PubMed^®, Cochrane Library^®, ScienceDirect^®, and Clinical Trials^®. After applying selection filters, 37 articles met the inclusion criteria. Data from these studies were extracted into a table, detailing authors, publication year, sample size, clinical trial phase, and endpoints, including ALSFRS-R scores, survival rates, and adverse events. Regarding survival, Edaravone demonstrated a significant survival benefit, with the active treatment groups living longer than the placebo groups. Other promising treatments, such as Sodium Phenylbutyrate and Taurursodiol, reduced mortality and hospitalizations, offering hope for improving patient outcomes. Tofersen also showed a survival increase, indicating substantial clinical potential. Safety profiles revealed common adverse effects, including headaches, dizziness, gastrointestinal issues, and increased fall risks, primarily affecting the nervous and digestive systems. These findings highlight the importance of careful risk–benefit evaluation in ALS treatment. Although the results indicate positive therapeutic developments, further trials are essential to confirm the long-term safety and efficacy of these treatments.

8.6. Pulmonary Mycobacterium Chimaera Intracellulare: A Rare Case Report

Brijesh Koyani ¹, Avani Mendpara ²

^1. 

Consultant Chest Physician at H J Doshi Hospital, Rajkot, Gujarat (India)

^2. 

Consultant Physician at H J Doshi Hospital, Rajkot, Gujarat (India)

Abstract: We report a rare case of pulmonary Mycobacterium chimaera intracellulare infection in a 69-year-old female with a history of pulmonary tuberculosis. The patient presented with chronic cough, weight loss, and shortness of breath and was successfully treated with a combination of antibiotics.

Introduction: Mycobacterium chimaera intracellulare is a rare and emerging pathogen that can cause pulmonary disease, particularly in immunocompromised individuals. We present a case of pulmonary Mycobacterium chimaera intracellulare infection in a 69-year-old female with a history of pulmonary tuberculosis.

Case Presentation: A 69-year-old female with a history of pulmonary tuberculosis presented to our hospital with a 3-month history of chronic cough, weight loss, and shortness of breath. Chest imaging revealed bilateral lung nodules and cavitations. Sputum samples were positive for Mycobacterium chimaera intracellulare, confirmed by molecular testing.

Treatment and Outcome: The patient was treated with a combination of antibiotics, including clarithromycin, rifampicin, and ethambutol. She showed significant clinical improvement, with the resolution of the symptoms and radiographic findings. The patient was treated for 12 months, with regular follow-up appointments to monitor her progress.

Discussion: This case highlights the importance of considering Mycobacterium chimaera intracellulare in the differential diagnosis of pulmonary infections, particularly in patients with a history of pulmonary tuberculosis. Early diagnosis and treatment are crucial to prevent disease progression and improve outcomes.

Conclusions: Pulmonary Mycobacterium chimaera intracellulare infection is a rare but important diagnosis to consider in patients with pulmonary symptoms and a history of tuberculosis. Prompt treatment with antibiotics can lead to significant clinical improvement and the resolution of symptoms.

8.7. The SAP–WASP–DGKα Axis in Rare Hematological Diseases

Elisa Gorla ^1,2, Luisa Racca ^1,2, Sara Centonze ^1,2, Sabrina Mula ³, Giulia Rossino ³, Valeria Malacarne ^1,3, Marcello Manfredi ^1,2, Davide Corà ^1,2, Alessandra Bertoni ¹, Andrea Graziani ⁴, Gianluca Baldanzi ¹

^1. 

Department of Translational Medicine, University of Piemonte Orientale, 28100 Novara, Italy

^2. 

Center for Translational Research on Allergic and Autoimmune Diseases (CAAD), University of Piemonte Orientale, 28100 Novara, Italy

^3. 

Department of Molecular Biotechnology and Health Sciences, Molecular Biotechnology Center (MBC), University of Turin, Turin, Italy

^4. 

Department of Molecular Biotechnology and Health Sciences, Molecular Biotechnology Center (MBC), University of Turin, Turin, Italy

Introduction: Wiskott—Aldrich Syndrome patients feature defects in T-cell activation and proliferation, as well as in megakaryopoiesis and platelets. We discovered that diacylglycerol kinase alpha (DGKα) constrains T-cell activation, and its activity is limited by a pathway comprising SAP (an adaptor mutated in Duncan’s disease) and the WAS protein (WASp). We propose that in the context of SAP or WASp deficiency, there is excessive DGKα activity contributing to diseases phenotypes.

Methods: In T-cells, we triggered immune synapse (IS) formation among Jurkat triple reporter cells and superantigen-loaded Raji cells. Jurkat triple reporter cells allow for the quantification of NFAT, NFκB, and AP-1 activity induction using flow cytometry. Similarly, we silenced WAS in the erythroleukemia cell line (HEL) and induced differentiation through PMA treatment, measuring cell elongation, spread, and CD41 induction.

Results: The IS analysis confirmed that SAP- and WASp-deficient cells have a reduced capacity to form IS and, surprisingly, DGKα inhibition further reduces it, underlining an important role of DGKα in shaping IS. The SAP- and WASp-deficient cells also showed a defect in the NFAT and AP-1 signaling pathways, whereas NFκB activity was unaffected. DGKα inhibition rescued both the NFAT and AP-1 defects but did not interfere with NFκB. In megakaryocytes, DGKα dampered thrombopoiesis and platelet aggregation. DGKα inhibition rescued the elongation defect of WASp-deficient HEL cells but was not able to restore CD41 induction.

Conclusions: Thus, in the absence of SAP or WASp, excessive DGKα activity consumes diacylglycerol, perturbing signaling. Indeed, DGKα inhibition is required for NFAT and AP-1 activation, and this can be compensated by DGK inhibitors. Similarly, excessive megakaryocyte DGKα activity in WAS reduces cytoskeletal remodeling, and this is also restored by DGK inhibitors. All of these observations suggest that inhibiting DGKα activity could represent a novel therapeutic approach for WAS.

9. Molecular Mechanisms of Neurodegeneration

9.1. Advancement in Liposomal Technology with Various Active Compounds for Neurodegenerative Disease Treatment

Elika Valehi, Gábor Katona, Dorina Gabriella Dobó, Ildikó Csóka

• University of Szeged, Department of Pharmaceutical Technology, Hungary

Neurodegenerative diseases, including Alzheimer’s, dementia, and Parkinson’s, pose significant global health challenges, leading to cognitive decline, motor impairment, and reduced quality of life. While conventional therapies can alleviate symptoms, they do not provide a cure. Therefore, there remains a need for more effective treatment strategies that can enhance drug bioavailability and therapeutic outcomes. A major challenge in treating these diseases is the blood–brain barrier (BBB), which restricts the passage of many central nervous system (CNS) active drugs, limiting their effectiveness. Intranasal drug delivery has emerged as a promising alternative, allowing drugs to bypass the BBB efficiently, ensuring rapid absorption while minimizing systemic side effects. Many CNS drugs suffer from poor solubility, stability, and permeability, reducing their therapeutic impact. Nanocarriers, specifically liposomes—nanoscale, lipid-based vesicles—offer a solution by protecting drugs from enzymatic degradation, enhancing absorption, and facilitating transport across the BBB. Liposomes provide advantages such as targeted drug delivery, controlled release, and improved bioavailability, making them ideal for CNS drug delivery via the nasal route. Our research focuses on developing and characterizing liposomal formulations for three CNS drugs, i.e., dopamine, vinpocetine, and donepezil, which suffer from low stability or bioavailability. By encapsulating these drugs in liposomes, we aim to enhance their solubility, stability, and BBB permeability, thereby improving their therapeutic potential. We focus on the impact of the liposome surface charge on critical factors such as mucoadhesion, drug release, and permeability, optimizing the drugs’ nasal applicability. We also evaluated their chemical and physical stability for long-term storage and clinical use. The results showed that we succeeded in developing robust, stable, and effective nasal liposomal drug delivery systems to improve treatment outcomes for neurodegenerative diseases. This research could lead to more efficient, noninvasive treatments, offering hope to millions worldwide.

Acknowledgments: Project No. TKP2021-EGA-32 was implemented with the support provided by the Ministry of Innovation and Technology of Hungary, financed under the TKP2021-EGA funding scheme.

9.2. Nanomechanics and Conformational Polymorphism in Neurotoxic Proteins: Understanding Amyloidogenic Neurodegenerative Diseases

Ishant Diwakar Dahake

• Department of Pharmaceutical Sciences, Rashtrasant Tukadoji Maharaj Nagpur University, Nagpur, 440033, India

Alzheimer’s, Parkinson’s, and Huntington’s are neurodegenerative diseases that are currently incurable and are uniquely defined by the deposition of neurotoxic proteins. Neurotoxic proteins are intrinsically disordered proteins with very high causality for neurodegeneration. Tremendous advances have been realized in molecular pathogenesis, with a view toward establishing causative genes and proteins. However, two important elements related to neurotoxic conformation and neuronal vulnerability remain unidentified. The prevailing hypothesis has been that pathogenic cascades are initiated by the conformational changes in the monomeric forms, and hence, these could be good targets for therapy. Single-molecule techniques, especially single-molecule force spectroscopy, have advanced the nanomechanics and conformational polymorphism of neurotoxic proteins. It has been shown that such polymorphism at the level of monomers is very strongly correlated with amyloidogenesis and neurotoxicity. Such polymorphism is, importantly, entirely absent in fibrillization-incompetent mutants but is enhanced by familial-disease mutations. There is also the case of pharmacological agents that inhibit β-conformational changes in monomers, dramatically reducing their polymorphism and associated neurotoxicity, indicating common molecular mechanisms for these diseases.

Advances in the manipulation and analysis of single molecules have dramatically enhanced our understanding of neurotoxic protein dynamics. Despite such advances, major questions remain about specific neurotoxic conformations and their role in selective neuronal degeneration. The solution to these challenges is crucial for the development of novel diagnostic, preventive, and therapeutic strategies against these devastating disorders, which carry significant social and clinical impacts.

This abstract encapsulates the emerging potential of single-molecule studies to pave the way for innovative solutions in managing amyloidogenic neurodegenerative diseases.

9.3. A Comprehensive Investigation into the Phytochemical Screening, Antimicrobial Sensitivity, and CNS-Depressant Properties of Piper Methysticum and Sansevieria Trifasciata

Md. Mizan ^1,2, Sahitto Biswas ²

^1. 

Department of Pharmacy, Dhaka International University, Satarkul, Badda, Dhaka-1212, Bangladesh

^2. 

Pharmacy Discipline, School of life Sciences, Khulna University, Khulna 9208, Bangladesh

Piper methysticum, traditionally used in the Pacific for religious, medical, and social purposes, has garnered recent fame as a nutraceutical for anxiety treatment. Sansevieria trifasciata (snake plant) has antibacterial, cytotoxic, and wound-healing properties. The ethanolic extracts of both plants were studied for their chemical group and their antimicrobial and CNS-depressant effects. The disc diffusion method was used to assess the antimicrobial activity of Piper methysticum and Sansevieria trifasciata. Their central nervous system (CNS)-depressant effects were evaluated using classical depression models, including hole cross, elevated maze, forced swimming, and tail suspension tests in Swiss albino mice. Mice were divided into control, positive control, and three test groups, with doses of 250 mg/kg and 500 mg/kg for the hole cross and elevated maze tests, and 50, 100, and 200 mg/kg for the forced swimming and tail suspension tests. Diazepam was used as the standard drug. The phytochemical screening of both plant extracts revealed the presence of alkaloids, flavonoids, reducing sugars, steroids, saponins, terpenoids, and amino acids. Both of the ethanolic extracts exhibited strong antibacterial activity in disk diffusion testing. S. trifasciata was most effective against E. coli and S. cerevisiae, while P. methysticum showed significant antimicrobial activity against Shigella dysenteriae and Salmonella typhi. In the hole cross test, both of the extracts significantly reduced locomotor activity across all doses (p < 0.01), indicating CNS-depressant effects. However, neither extract showed significant changes in the number of entries or the time spent in the open arms during the elevated maze test. Both of the extracts significantly reduced the immobility time in the forced swimming and tail suspension tests (p < 0.01 and p < 0.001), with diazepam increasing the immobility time (p < 0.001). These results suggest that both of the plant extracts have CNS-depressive and antidepressant-like effects. Both of the ethanolic extracts of P. methysticum and S. trifasciata demonstrate antimicrobial and CNS-depressant effects. Further studies are needed to continue pharmacological research for their potential use in medicine.

9.4. Coumarin–Benzodiazepine Hybrid Modulators of AMPA Receptors: From Subunit Selectivity Toward Their Next-Stage Potential in a Fully Integrated Neuronal Environment

Mohammad Bdair, Mohammad Qneibi

• Department of Biomedical Sciences, Faculty of Medicine and Health Sciences, An-Najah National University, Nablus, Palestine

The dysregulation of α-amino-3-hydroxy-5-methyl-4-isoxazole propionic acid (AMPA) receptors underlies a variety of neurological conditions, including epilepsy and neurodegenerative disorders. Pharmacological approaches that selectively modulate AMPA receptor subunits offer significant promise in mitigating glutamate-mediated excitotoxicity. We investigated eight newly synthesized coumarin–benzodiazepine (CD) hybrid derivatives designed as negative allosteric modulators of AMPA receptors, seeking both potency and selectivity. Using whole-cell patch-clamp electrophysiology in HEK293 cells expressing specific AMPA receptor subunits (GluA1–4), we evaluated the ability of each compound to alter receptor kinetics, particularly desensitization and deactivation. Among the tested derivatives, one molecule (CD8) exhibited a notably high affinity for AMPA receptors, with potent inhibition observed across multiple subunit combinations. Crucially, CD8 displayed minimal off-target activity against kainate and NMDA receptors, underscoring its selectivity for AMPA receptor subtypes. Structurally, CD8’s carboxyl substituent at the para position of the phenyl ring emerged as pivotal for enhanced receptor binding and negative modulation. Furthermore, kinetics analyses revealed that certain derivatives, including CD8, not only attenuated the peak current amplitude but also accelerated deactivation rates and reduced desensitization—indicative of a robust negative allosteric mechanism that is distinct from the orthosteric glutamate-binding site. Taken together, these findings highlight the potential of coumarin–benzodiazepine hybrids as selective modulators of AMPA receptors. By dampening hyperexcitatory neuronal signaling through targeted receptor inhibition, these derivatives hold promise as a therapeutic avenue for a range of central nervous system pathologies. Future research will probe how these compounds interact with auxiliary proteins (e.g., TARPs) to further optimize AMPA receptor gating in native neuronal environments, thereby refining their translational potential for neurological conditions associated with abnormal glutamatergic transmission.

9.5. Design of Partial Agonists of ADAMTS Metalloproteinases as Therapeutics for Neurodegenerative Diseases

David Joshua Ferguson

• Adjunct Faculty, Department of Chemistry, Virginia Peninsula Community College, Hampton, Virginia, 23666, United States of America

Neurodegenerative diseases (NDDs), including Alzheimer’s disease, Parkinson’s disease, and multiple sclerosis, are progressive disorders characterized by neuronal dysfunction and structural instability. Central to their pathology are aberrations in extracellular matrix (ECM) remodeling and chronic neuroinflammation, processes regulated by the ADAMTS family of metalloproteinases. The dysregulated activity of specific ADAMTS isoforms, such as ADAMTS-4 and ADAMTS-5, contributes to pathological ECM degradation, triggering neuroinflammatory cascades, synaptic disruption, and neuronal loss. This article proposes a novel therapeutic approach: the development of partial agonists targeting ADAMTS enzymes. Unlike broad-spectrum inhibitors, which can suppress essential protease functions, partial agonists offer a balanced strategy by selectively modulating enzymatic activity. These agents aim to mitigate pathological ECM degradation while preserving physiological ECM dynamics, addressing both neuroinflammation and ECM imbalances.

The design of partial agonists leverages bioisosteric principles to create isoform-specific targeting agents with high therapeutic precision. Structural scaffolds inspired by protease–substrate interactions are central to this approach. Additionally, innovative drug delivery systems, including nanoparticle encapsulation and prodrug strategies, are explored to overcome the formidable challenge of crossing the blood–brain barrier and ensure sustained drug release in the central nervous system. Advanced delivery platforms enhance therapeutic efficacy while minimizing systemic side effects, which is a critical consideration for chronic neurodegenerative disease management and therapy and developing potential curative strategies.

This research highlights the potential of ADAMTS-targeting partial agonists to transform NDD therapeutics by restoring ECM integrity and reducing neuroinflammation. Through these mechanisms, partial agonists can enhance neuronal resilience, stabilize synaptic networks, and significantly improve clinical outcomes, offering new hope for individuals affected by debilitating neurodegenerative conditions.

9.6. The Neuroprotective Effects of the Flowers of Camellia japonica var. Hagoromo: A Potential Candidate for Neurodegenerative Disease Treatment

Ezgi Nur YUKSEK ¹, A. Gonzalez Pereira ¹, Maria Carpena Rodríguez ¹, Aurora Silva ², Clara Grosso ², M. Fátima Barroso ², Miguel Angel Prieto ³

^1. 

Universidade de Vigo, Nutrition and Bromatology Group, Department of Analytical Chemistry and Food Science, Instituto de Agroecoloxía e Alimentación (IAA)—CITEXVI, 36310 Vigo, Spain

^2. 

REQUIMTE/LAQV, Instituto Superior de Engenharia do Porto, Instituto Politécnico do Porto, Rua Dr. António Bernardino de Almeida 431, 4249-015 Porto, Portugal

^3. 

Universidade de Vigo, Nutrition and Bromatology Group, Department of Analytical Chemistry and Food Science, Instituto de Agroecoloxía e Alimentación (IAA)—CITEXVI, 36310 Vigo, Spain

Neurodegenerative diseases, such as Alzheimer’s and Parkinson’s, are characterized by cholinergic dysfunction and oxidative stress, leading to progressive cognitive decline. The lack of effective treatments for these disorders makes the search for naturally derived neuroprotective agents increasingly important. Various Camellia species have demonstrated neuroprotective effects, primarily attributed to their rich phytochemical composition. However, no previous study has investigated the neuroprotective properties of Camellia japonica, particularly the Hagoromo cultivar. This study evaluates the neuroprotective potential of C. japonica var. Hagoromo flower extract by evaluating its inhibitory effects on acetylcholinesterase (AChE) and butyrylcholinesterase (BuChE) enzymes using previously established methods. The extract showed a dose-dependent increase in AChE inhibition, with 1 mg/mL achieving 7% inhibition and 2 mg/mL reaching 22%. For BuChE inhibition, the extract showed its highest activity at 1 mg/mL, reaching 37% inhibition. These findings indicate potential neuroprotective properties, although the observed inhibition levels were lower than the levels of well-known neuroprotective compounds, such as fucoxanthin and polyphenols found in seaweeds [8,9]. However, this study was conducted using unpurified extracts, which may limit the accuracy of the observed effects. Therefore, further studies should focus on purifying and isolating the bioactive compounds present in the sample to provide a more precise assessment of their neuroprotective potential. Previous studies suggest that catechins, quercetin, kaempferol, and their derivatives are the primary neuroprotective phytochemicals in Camellia. These compounds exert their effects by synergistically enhancing endogenous antioxidant defenses, regulating neurotrophic signaling pathways, and reducing neuroinflammation [10]. Future research should optimize extraction parameters, identify key active compounds, and conduct in vivo evaluations to further validate the therapeutic potential of C. japonica in neurodegenerative disorders.

9.7. Protein Aggregating and Misfolding in Neurodegenerative Diseases: A Meta-Analysis

Mohini Kalra ¹, Tamanna Kalra ²

^1. 

Amity Institute of Pharmacy, Amity University Haryana, Gurugram (Manesar), Haryana 122413, India

^2. 

Department of Psychology, Lakshmibai College, University of Delhi, Delhi 110052, India

Overview: A global health concern is neurodegenerative illnesses, which are brought on by an increasing loss of the neurons, one of the main causes of cognitive and physical deterioration in these illnesses. The death of brain cells or nervous system cells is the cause of this neurological loss. Huntington’s disease, Parkinson’s disease, and Alzheimer’s disease are among the most common conditions that share protein misfolding and aggregation. According to a WHO news report, neurological disorders impact more than one in three people and are a leading cause of disability worldwide. Since 1990, the DALYs (Disability-Adjusted Life Years) for these illnesses have gone up by 18%. The prevalence of Alzheimer’s disease is estimated to be 55 million people worldwide, and for Parkinson’s disease, it is 10 million globally.

Objective: We wished to achieve the following:

• To review and research the role of protein misfolding and aggregation in neurodegenerative diseases.
• To find common molecular mechanisms for neurological disorders and the target sites for a therapeutic effect.
• To evaluate the effect of genetic, environmental, and lifestyle choices on protein misfolding.

Methods: A thorough search of the literature was conducted in several large medical databases. This study was carried out in the form of a meta-analysis of the available literature. Key findings on protein misfolding and aggregation, the participant characteristics, and the study design were all included in the data extraction process. The data analysis was performed, and the results were presented.

Results: We determined common pathways and sites for protein misfolding in neurodegenerative disorders. The results reflect upon the different factors affecting the aggregation and misfolding of proteins, and using them, the target sites for a therapeutic effect can be confirmed.

9.8. Study of Hematopoiesis in Brain Trauma: Exploring New Approaches to Regulating Neuroinflammation and Neurogenesis

Alexander Dygai, Edgar Pan, Natalia Ermakova, Mariia Zhukova, Valentina Pan, Irina Zharkikh, Victoria Skurikhina

• Institute of General Pathology and Pathophysiology, 125315 Moscow, Russia

Background: Current treatments for traumatic brain injury (TBI) commonly focus on symptom management and lack strategies to prevent or delay the development of neurodegenerative processes. The lack of understanding of TBI mechanisms hinders the development of effective and safe approaches for anti-inflammatory therapy and the stimulation of regeneration in damaged and/or lost neurons. The study of the blood system in TBI may provide new biomarkers for the development of neurodegenerative processes and methods for regulating neuroinflammation and neuroregeneration.

Materials and Methods: Male ICR mice, 12–14 weeks old, were used in this experiment. Traumatic brain injury was induced by focal impact using a weight-drop model [11]. The neurological status of the mice was assessed before the injury and at 6 h post-injury, as well as on days 1, 3, 7, 14, 21, and 42 post-injury. Histological examination (hematoxylin and eosin staining), immunohistochemical staining, and an analysis of the expression of astrocyte, mature, and immature neuron markers in the brain were performed. The content of hematopoietic cells in the blood and bone marrow was studied via staining, using the May–Grünwald–Giemsa staining method, along with the quantitative and qualitative composition of hematopoietic niches [12]. Additionally, an in vitro analysis of hematopoietic and progenitor cells was performed. The sympatholytic drug reserpine was used to modulate neuroinflammation and neurogenesis.

Results: Reserpine, via decreasing hematopoietic activity, reduced brain damage, prevented neuroinflammation, and facilitated neurogenesis in mice subjected to TBI.

Conclusions: A pharmacological blockade of the sympathetic component of hematopoiesis regulation may serve as a basis for developing new approaches to reduce neuroinflammation and promote neurogenesis in TBIs.

9.9. The Impact of Persistent Neurological Manifestations in Long COVID-19 Syndrome on Daily Life Quality

Roxana Gheorghita ¹, Iuliana Soldanescu ², Otilia Cramariuc ², Olga Caliman Sturdza ³

^1. 

Faculty of Medicine and Biological Sciences, Stefan cel Mare University of Suceava, Romania

^2. 

Stefan cel Mare University of Suceava, Romania

^3. 

Faculty of Medicine and Biological Sciences, Stefan cel Mare University of Suceava, Romania Suceava Emergency County Clinical Hospital

Long COVID (L-C19) leads to significant consequences and affects the quality of life of patients and their families as well. The most common neurological symptoms are headaches, brain fog, sleep disturbances, chronic fatigue, forgetfulness, and even more serious conditions, such as balance disorders, anxiety, and depression. Headache is the earliest and most common symptom of L-C19 and is sometimes accompanied by vomiting or nausea. Patients often report difficulties with concentration and learning and a reduced efficiency in daily activities. Even though these disorders may be caused by an acute illness, biochemical changes caused by SARS-CoV-2 in the nervous system can be an important factor. L-C19 also affects the peripheral nerves, leading to paresthesia, muscle aches, and weakness throughout the body. The duration of these symptoms is often unknown, and the treatment is predominantly symptom-based and difficult to predict. Other dysfunctions may include dysautonomia, a condition affecting the autonomic nervous system, responsible for regulating automatic processes, such as heart rate, blood pressure, and digestion. This disruption may lead to symptoms like dizziness and fatigue, is often accompanied by neuropsychiatric challenges, and can alter cerebral blood flow, contributing to light-headedness, trouble focusing, and emotional instability. Furthermore, the SARS-CoV-2 virus may disrupt mitochondrial function, an important component of cellular energy production. Mitochondrial dysfunction can manifest as fatigue, cognitive challenges, and mood disorders, given the brain’s sensitivity to shifts in energy metabolism.

Acknowledgments: This work was supported by a grant of the Ministry of Research, Innovation and Digitization, under the Romania’s National Recovery and Resilience Plan Funded by the EU “Next Generation EU” program, project “Artificial intelligence-powered personalized health and genomics libraries for the analysis of long-term effects in COVID-19 patients (AI-PHGL-COVID)”, number 760073/23.05.2023, code 285/30.11.2022, within Pillar III, Component C9, Investment 8.

9.10. A Study of the Neuroprotective Therapeutic Effects of Rubus Fruticosus Extracts Through the Modulation of Cognitive Deficits Specific to Alzheimer’s Disease Pathology

Paula Denisa Saragea, Lucian Hrițcu

• Biology Department, “Alexandru Ioan Cuza” University, Iași, Romania

As one of the principal etiological determinants of irreversible progressive dementia, the pathogenesis of Alzheimer’s disease (AD) is predominantly ascribed to the intracellular accumulation of neurofibrillary aggregates composed of hyperphosphorylated tau proteins within limbic and cortical regions, alongside the extracellular deposition of dense/diffuse β-amyloid neuritic plaques. AD represents a multifactorial, conformational, neurodegenerative disorder with a predominant proteinopathic character, exhibiting an insidious onset and an irreversible progression.

The selected experimental model, the 5xFAD mouse, exhibits severe amyloidosis, harboring five pathogenic mutations within the human transgenes APP (amyloid precursor protein) and PSEN1 (presenilin-1), leading to a fulminant disease progression. The experimental cohorts comprised a negative control, a positive control treated with galantamine (3 mg/kg), and two groups per tested extract, corresponding to the two predefined concentrations (50 mg/kg and 100 mg/kg). The impact on cognitive performance was assessed through a battery of behavioral tests: a Y-maze test (YMT), open-field test (OFT), novel object recognition test (NORT), elevated plus maze test (EPM), forced swimming test (FST), and radial arm maze test (RAM). The pharmacokinetic profiles of the investigated compounds were analyzed in silico using computational tools, such as SwissADME and pKCSM.

The study highlights the neuroprotective potential of Rubus fruticosus extracts, demonstrating a significant enhancement in cognitive performance and exploratory activity across the behavioral tests, alongside a marked reduction in stress levels, indicative of anxiolytic and antidepressant properties. In silico analyses revealed favorable pharmacokinetic attributes conducive to ameliorating memory deficits, including the capacity of the compounds to cross the blood–brain barrier, promising interactions with key molecular targets, and a bioactivity profile suggestive of drug-like potential.

The findings suggest that Rubus fruticosus extracts exhibit promising pharmacokinetic profiles and neuroprotective pharmacological potential, underscoring the imperative for further multifaceted, multidisciplinary investigations to validate their underlying mechanisms and clinical applicability.