World Academy of Science, Engineering and Technology

Authors: Sidra Ghori, Muhammad Kashan Surahio, Ehsan Ali Shaikh

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Poor diet, physical inactivity, tobacco use, and alcohol consumption are well-established risk factors for chronic diseases and premature mortality. These behaviors are particularly concerning among university students and are often linked to psychological distress, especially in academic programs involving practicum components, such as nursing and teaching. Understanding how these risk behaviors cluster and relate to psychological well-being and coping mechanisms is essential for effective health promotion. This study was conducted at Indus Medical College, involving a total of 750 undergraduate students enrolled in nursing, midwifery, and teacher education programs. A comprehensive survey approach was used, incorporating the Lifestyle Behaviour Questionnaire (LBQ), the General Health Questionnaire (GHQ), and the Ways of Coping Questionnaire (WCQ) to assess lifestyle practices, psychological distress, and coping strategies. The results indicated a high prevalence of health-risk behaviors among participants, including alcohol consumption (93.2%), unhealthy diet (26.3%), physical inactivity (26%), tobacco smoking (17%), cannabis use (11.6%), and significant levels of stress (41.9%). Based on behavioral patterns, students clustered into two main groups: those engaging in risk behaviors and those practicing positive health behaviors. Students in the risk-behavior group reported higher psychological distress and primarily used passive coping strategies such as escape-avoidance. These findings are concerning not only for the current health and academic success of these students but also for their future roles as health professionals and educators. The results highlight the need for comprehensive, university-based health promotion initiatives aimed at reducing multiple risk behaviors and supporting student well-being, particularly in institutions like Indus Medical College.

Keywords: psychological distress, lifestyle behaviours, health promotion, undergraduate students, Indus Medical College, coping strategies, nursing education, teacher education, risk behaviors, student well-being

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Authors: Leonel Vázquez Carrasco, Jesús Antonio Guzmán Doroteo, Víctor Tonatiuh Acosta Mendoza

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This paper presents the validation of a hybrid motor-assessment system that integrates EEG (Unicorn Hybrid Black) and EMG (BIOPAC MP36) signals during a joystick task built in Unity, with data streams synchronized via Lab Streaming Layer (LSL). The protocol comprised baseline phases (pre and post), a practice phase, and eight blocks of 40 trials each. The study involved six participants from two age groups, young adults and older adults. All signals were synchronized and processed in Python, where inter-group differences were observed through the resulting plots and metrics. The incorporation of transcranial electrical stimulation (TES) is envisioned for future versions.

Keywords: motor performance evaluation, EEG, EMG, kinematic

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Authors: Aikaterini Myrto Koliousi, Maria Oikonomou, Athanassios Mihailidis

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The temporomandibular joint (TMJ) is a complex anatomical and bio-mechanical structure critical to mastication, speech, and overall cranio-facial function. It is also vulnerable to dysfunctions resulting from injuries sustained by cyclists and motorcyclists following falls and traffic accidents. This review consolidates finite element analyses of the temporoman-dibular joint published in the last fifteen years. The content of over one hundred studies is roughly divided into three topics: orthodontic treat-ment, which reshapes pressure and tension in the area; craniofacial deformities—chiefly mandibular projection; and joint dysfunctions, in which temporary or permanent disc displacement and disc perforation significantly redefine contact pressures. Together, these models clarify how mechanical responses differ with treatment, anatomy, and pathol-ogy. The study also reviews treatment approaches, including occlusal splints and prosthetic joint replacements, in the context of biomechani-cal insight provided by finite element (FE) analysis. Prompted by the frequent occurrence of TMJ-related injuries among cyclists and motorcyclists, this study presents a FE analysis of the joint to reproduce its mechanical response during a mild lateral tip-over, typical of everyday riding. The FE model was constructed with materi-al properties taken from the literature, and the simulation captures the mechanical quantities related to TMJ injury in such falls. This work contributes to the understanding of TMJ biomechanics and supports the design of more effective diagnostic and therapeutic inter-ventions.

Keywords: biomechanical modeling, TMJ, TMJ injuries, TMJ trauma

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Authors: Vasileios E. Katsigiannis, Stavros T. Miloulis, Ioannis Kakkos, Ioannis A. Vezakis, Naomi C. Buntsma, Joyce Rops, Edwin van der Pol, George K. Matsopoulos

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Stroke is a leading cause of mortality and long-term disability worldwide, with swift and accurate diagnosis being vital for ensuring timely therapeutic intervention and improved clinical outcomes. Although neuroimaging techniques, such as computed tomography (CT) and magnetic resonance imaging (MRI), are included in the current standard of care, availability limitations across clinical sites and inherent processing delays can impede the rapid initiation of appropriate treatment. Recent advances suggest that blood-based biomarkers, such as extracellular vesicles (EVs), offer a promising, minimally invasive alternative to neuroimaging techniques by reflecting the underlying pathophysiological changes associated with different stroke subtypes. However, very few studies have quantified the capacity of EVs as biomarkers for diagnosis and monitoring. Therefore, in this study, we developed an AI-driven diagnostic framework that integrates EV-derived features with machine learning (ML) algorithms to distinguish between hemorrhagic stroke (HS), ischemic stroke due to small vessel occlusion (ISSVO), and ischemic stroke due to large vessel occlusion (ISLVO). The study incorporated a dataset of blood plasma samples from 123 stroke patients, where flow cytometry was used to identify EVs and quantify features such as size distribution metrics, concentration, and emitted fluorescence intensity. These features were analyzed using multiple ML classifiers—including Gradient Boosting, eXtreme Gradient Boosting, k-nearest neighbors, and Logistic Regression—with the latter achieving a weighted accuracy of 63% and an F1-score of 61%. Findings highlight the potential of an AI-enhanced EV biomarker approach complementing traditional imaging to streamline stroke subtype diagnosis in healthcare settings and improve clinical workflows.

Keywords: biomarkers, decision support system, extracellular vesicles, hemorrhagic stroke, ischemic stroke, machine learning

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Authors: Sara Bigagli, Giacomo Falaschi

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Clearaligner therapy has gained widespread acceptance as a comfortable and aesthetic alternative to fixed orthodontic appliances. While previous research has focused on the biomechanical and periodontal implications of aligner design, particularly the position of the gingival margin, there is limited evidence addressing patient-reported soft tissue discomfort. This gap is especially relevant in cases involving composite attachments, which may influence how the vestibular edge of the aligner interacts with the surrounding soft tissues. Poor adaptation or overextensions of the aligner margin can result in irritation, soreness, and inner lip discomfort. This study highlights the need for a deeper understanding of soft tissue responses to aligner design, with the goal of optimizing both clinical outcomes and patient comfort. The objective of the study is to investigate the relationship between aligners fit(Invisalign,Align technology), and soft tissue discomfort in cases where composite attachments interfere with complete aligner seating. This pilot study included six patients undergoing aligner therapy.For all of them the treatment plan provided aligners with composite attachments. In all patients, the brackets and therefore the respective aligners were applied and finally the images were acquired by iTero scanner. The images thus ob-tained were analyzed using OrthoCAD software (v. 5.9.1.50) in open shell view (arches oriented in occlusion) measuring the distance between the gingival margin (GV) of all teeth with attachments and the vestibular edge of the aligner (VE). One week after the implantation, a clinical follow-up of the patients was performed, asking them to declare discomfort using a numerical scale of 1-10. Considering all the measure-ments made (36 measurements on teeth with attachments) the GV-VE distance was found to be between a minimum of 0.3mm and a maximum of 2.2mm, with an average value of 0.8mm For patients who reported soft tissue discomfort great-er than or equal to 6 (n=3), the GV-VE distance ranged from 1.2mm to 2.2mm with a mean value of 1.4mm. In the three patients with discomfort of inner lips and cheeks, the presence of composite attachments altered the seating of the aligner, leading to a detectable gap at the gingival edge, particularly in the buccal anterior region. In pilot study it appears that patients with discomfort averaged greater gaps between the gingival margin and the vestibular edge of the aligner; this suggests that accurate assessment of aligner fit and angulation in areas with attachment has an important role in the design protocols and edge contouring during aligner fabrication. Further studies are needed to clarify the threshold that can lead to discomfort for the patient.

Keywords: attachement, aesthetic, aligner, soft tissue, discomfort

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Authors: S. A. Mukhametshin, E. M. Gilyazova, A. Kh. Valiullina, E. R. Bulatov

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CAR-T cell-based adoptive immunotherapy has demonstrated high efficiency in the treatment of a range of hematological malignancies. However, autologous CAR-T cells lose cytotoxicity in a suppressive tumor microenvironment. Thus, approaches to developing universal allogeneic CAR-T cell products are being advanced. The major limitations of such therapy are graft-versus-host disease and graft rejection. Improvement of the safety of allogeneic CAR-T therapy can be achieved by knocking out genes responsible for these side effects: the TRAC gene (encoding the α-subunit of the T-cell receptor) and the B2M gene (beta-2-microglobulin, a component of the major histocompatibility complex class I). Design of guide RNA (gRNA) for TRAC and B2M genes with the CRISPR/Cas9 system was performed through Knockout Guide Designer software (Synthego, USA), considering high target affinity and minimizing off-target sites. The designed gRNAs were ligated into pSpCas9(BB)-2A-GFP (PX458) vector and transformed into the Escherichia coli DH5α strain. The plasmids were verified by Sanger sequencing. HEK293FT cells were transfected by using the plasmids with polyethylenimine (PEI ‘Max,’ Polysciences, Germany). The GFP expression was quantified after 24 hours using a Nexcope NIB910-FL fluorescence microscope (Nexcope, China). The percentage of GFP-positive cells was quantified by a BD FACSAria III flow cytometer (BD Biosciences, USA). Knockout efficiency was quantified using real-time PCR. Peripheral blood mononuclear cells were isolated by Ficoll gradient centrifugation. T-lymphocytes were activated for 72 hours by T cell TransAct (Miltenyi, USA) in the presence of IL-2 (SciStore, Russia). PX458 plasmid containing gRNA template of the TRAC gene was electroporated into T-cells by Neon™ NxT Electroporation System (Thermo Fisher Scientific, USA). The knockout efficiency of the TRAC gene was analyzed 5 days later by flow cytometry using APC anti-human CD3 antibody (HIT3a, BioLegend, USA). Cells were lysed with RIPA buffer (Bio-Rad, USA). Proteins were separated using SDS-PAGE electrophoresis and transferred to a PVDF membrane. The membrane was stained with HRP-conjugated β-actin antibodies (2D1D10, GenScript Biotech, USA), primary antibodies anti-CD3 (ab699, Abcam, USA), anti-PTCRA (a7150, ABclonal, Germany), and secondary antibodies Goat anti-mouse IgG (ab205719, Abcam, USA) and Goat anti-rabbit IgG (ab97051, Abcam, USA), respectively. Protein level was normalized to β-actin. During CRISPR/Cas9 gRNA design, two sequences were obtained: for the TRAC gene — 5' GCUGGUACACGGCAGGGUCA 3', and for the B2M gene — 5' AAGUCAACUUCAAUGUCGGA 3'. Transfection of HEK293FT cell line yielded 37.9% and 30.0% GFP-positive cells after TRAC and B2M knockout, respectively, following a reduction in target gene expression by 31.0% (p<0.05) and 40.4% (p<0.05). A decrease in TCR-CD3 complex expression in T-lymphocytes was observed at 26.1% (low intensity, 1500V/13ms/3 pulses), 24.12% (medium intensity, 1500V/17ms/2 pulses), and 26.3% (high intensity, 1550V/13ms/3 pulses). Protein levels of PTCRA were decreased by 23.2%, 27.2%, and 29.7%, respectively. The obtained results will be used to transition toward generating universal allogeneic CAR-T lymphocytes with knockout of genes responsible for alloreactivity and rejection syndrome. The next stages of research include B2M knockout in T-lymphocytes and subsequent testing of donor-derived CAR-T cells for reactivity in co-culture with HLA-mismatched donor cells. Additionally, cytotoxicity assays will be performed in co-culture with tumor cell lines, comparing the effects to non-genome-edited CAR-T lymphocytes.

Keywords: allogeneic CAR-T therapy, CRISPR/Cas9, cancer therapy, adoptive cell therapy

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Authors: Marc T. Morris, Pallav Joshi, Georgina H. Joyce, Samuel J. Davis, Ulban Adhikary, Mikael Boden, Gerhard Schenk

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Antimicrobial resistance (AMR) is arguably the greatest threat faced by global healthcare. However, resistance to β-lactam antibiotics, and the last-resort carbapenems especially, is continually spreading in clinical settings. The spread of metallo-β-lactamases (MBLs) are of particular concern, as there are no inhibitors currently available for clinical use, as opposed to several clinically useful serine-β-lactamase inhibitors (e.g. clavulanic acid, sulbactam). Within MBLs, the diversity and clinical abundance of the B1 and B2 MBL subgroups, which include globally distributed members such as NDM-1 or CphA, are well-understood and characterized, but this is not the case for the more divergent, usually environmental B3 MBL subgroup. Indeed, the B3 subgroup displays much broader active site & sequence diversity than other β-lactamases, as well as a concerning frequency of promiscuous β-lactamase activity in non-antibiotic-degrading members across the breadth of the family. In addition, extremely efficient B3 β-lactamases such as AIM-1, the most-efficient known MBL, are now appearing in clinical environments and represent a major threat to global health. The B3 MBLs thus represents a high-risk reservoir of potentially novel vectors of antimicrobial resistance, while simultaneously being the least studied and understood subgroup. Using a combination of phylogenetic analyses and in vitro kinetic characterization, we provide a clearer-than-ever picture of the diverse catalytic landscape of B3 MBLs, with the purpose of establishing the risk profile for this MBL subgroup. In particular, we connect changes in β-lactamase activity with key active site residues, which we have previously shown to drastically affect the catalytic properties of proteins from the MBL-fold superfamily. Indeed, we have previously shown that key mutations in viral anti-Pycsar proteins and some key B3 MBLs allow the up- or down-regulation of β-lactamase activity with a concurrent opposite decrease or increase in nuclease activity for example. Using ancestral sequence reconstruction, we describe how β-lactamase activity evolved in B3 MBLs, which evolved distinctly to that of B1 and B2 MBLs, and correlate β-lactamase catalysis with key distinctive structural features of B3 MBLs.

Keywords: antimicrobial resistance, carbapenemase, ancestral sequence reconstruction, catalysis, metallo-β-lactamase

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Authors: Iman Khazrak, Shakhnoza Takhirova, Mostafa M. Rezaee, Mehrdad Yadollahi, Robert C. Green II, Shuteng Niu

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The development of accurate medical image classification models is often constrained by privacy concerns and data scarcity for certain conditions, leading to small and imbalanced datasets. To address these limitations, this study explores the use of generative models, such as Denoising Diffusion Probabilistic Models (DDPM) and Progressive Growing Generative Adversarial Networks (PGGANs), for dataset augmentation. The research introduces a framework to assess the impact of synthetic images generated by DDPM and PGGANs on the performance of four models: a custom CNN, Untrained VGG16, Pretrained VGG16, and Pretrained ResNet50. Experiments were conducted using Random Sampling and Greedy K Sampling to create small, imbalanced datasets. The synthetic images were evaluated using Frechet Inception Distance (FID) and compared to original datasets through classification metrics. The results show that DDPM consistently generated more realistic images with lower FID scores and significantly outperformed PGGANs in improving classification metrics across all models and datasets. Incorporating DDPM-generated images into the original datasets increased accuracy by up to 6%, enhancing model robustness and stability, particularly in imbalanced scenarios. Random Sampling demonstrated superior stability, while Greedy K Sampling offered diversity at the cost of higher FID scores. This study highlights the efficacy of DDPM in augmenting small, imbalanced medical image datasets, improving model performance by balancing the dataset and expanding its size.

Keywords: medical image augmentation, generative models, progressive growing generative adversarial networks, denoising diffusion probabilistic models, synthetic data integration

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Authors: Hicham Loumissi, Adil Barra, Najat Messaoudi, Othmane Elbadlaoui , Bahloul Bensassi, Hicham Medromi

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Photoplethysmography (PPG) is susceptible to noise, affecting vital sign monitoring accuracy. Traditional hardware filters offer simplicity but lack adaptability. Software filtering provides flexibility but can introduce processing delays and computational overhead. This paper investigates a hybrid filtering approach combining traditional RC filters with Arduino IDE-based software filtering for the MAX30102 sensor. The core challenge is to optimize signal quality while minimizing the drawbacks of each method. An RC filter reduces high-frequency noise and a moving average filter implemented on an Arduino Mega refines the signal. Data was collected in controlled settings. Performance was evaluated based on SNR, heart rate accuracy, and computational load. Results demonstrate the hybrid approach enhances PPG signal quality with improved noise reduction and adaptability, with acceptable computational load. This solution combines the simplicity of hardware filters with the adaptability of software filters, offering a robust solution for wearable health monitoring devices.

Keywords: photoplethysmography , hybrid filtering, heart rate monitoring, signal processing, max30102.

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Authors: Ananta Raj Adhikari

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The use of polymer composites has seen a remarkable rise in recent years, driving progress across multiple areas of material science. Polymers often lack the necessary surface properties required for certain applications. However, surface modification techniques have become essential in the plastics and various other industries, enabling the transformation of low-cost materials into high-value end products. While numerous advanced technologies have been developed to modify these materials, there remains a demand for methods that are simple, energy-efficient, controllable, cost-effective, timesaving, and environmentally sustainable. Electromagnetic wave can significantly modify materials both positively and negatively depending on their applications. Among these, microwave technology stands out as a powerful approach for material synthesis and modification. Its advantages - such as rapid, selective, and uniform heating, along with direct molecular-level energy transfer - make it particularly promising. This presentation will highlight the use of microwave energy as an alternative method for processing materials, with a focus on current research in our lab, especially its potential applications in the medical field.

Keywords: material processing, microwave radiation, polymer composites, surface modification

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Authors: Blake Suzelis

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Miracles, magic, and the supernatural are all seen as extraordinary; however, these experiences may have some knowable origin that is presently unknown. Spontaneous cancer remission,the disappearance of cancer lacking what allopathic medicine would consider adequate treatment, is a poorly understood yet incredible phenomenon that, if better understood, offers hope that cancer could be seen as a teacher and not a death sentence. This paper aimed to deepen understanding of the mind-body-spirit role in spontaneous cancer remission from a holism, as opposed to scientific materialism, theoretical approach. The research question asked was, “What is the correlation between the path to spontaneous cancer remission and the journey to spiritual enlightenment as narrated in The Hero’s Journey and depicted in the archetypes of the tarot Major Arcana?” To identify a process for spontaneous cancer remission, this first holism quantitative survey study of cancer utilized an online self-administered special-purpose SurveyMonkey survey. Univariate and bivariate data analysis, along with descriptive statistics, were used to analyze the sixteen eligible participant surveys. A three-phase path emerged from the results involving (a) seeking answers externally while learning to calm the survival brain, (b) using the thinking brain to take responsibility for the body through doing activities, and (c) making choices for one’s unique path by learning to trust the intuitive heart through self-forgiveness, self-acceptance, and self-love. Further, thisvital evolution required a growth mindset and surrendering expectancy. The significance of the results aids in supporting allopathic medical approaches for acute care, while cancer and chronic illnesses may be best approached through a holism theoretical approach. Furthermore, the research findings highlight the connection between a quantum physics understanding of reality and what we refer to as spirituality, thereby bridging the gap between science and spirituality. Future studies may continue to build the process toward spontaneous cancer remission and our understanding of reality.

Keywords: mind-body-spirit, spontaneous cancer remission, mindbody medicine, holism, quantum physics

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Authors: Munsa Gill Kang

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Coeliac disease (CD) is a chronic autoimmune enteropathy triggered by ingestion of gluten in genetically susceptible individuals, resulting in intestinal inflammation, villous atrophy, and malabsorption. The cornerstone of treatment remains lifelong adherence to a strict gluten-free diet (GFD), which effectively controls symptoms and promotes mucosal healing in most patients. However, adherence challenges, inadvertent gluten exposure, and persistent symptoms in a subset of patients highlight the unmet need for additional therapeutic options. Over the past decade, advances in understanding the immunopathogenesis and molecular mechanisms underlying CD have fueled the development of novel therapies aimed at supplementing or potentially replacing the GFD. This review critically examines emerging treatment modalities targeting various pathogenic steps in CD. Enzymatic therapies such as latiglutenase and TAK-062 are designed to degrade immunogenic gluten peptides in the gastrointestinal tract, reducing their capacity to trigger an immune response. Zonulin antagonists like larazotide acetate work by restoring intestinal barrier integrity and preventing gluten peptide translocation, thus mitigating mucosal immune activation. Immunomodulatory strategies include peptide-based vaccines and immunotherapies that seek to induce specific immune tolerance to gluten, although some candidates have encountered challenges in clinical trials. Biologic agents targeting cytokines such as interleukin-15 (IL-15) have shown promise, especially for refractory CD where immune dysregulation persists despite strict GFD adherence. Moreover, modulation of the gut microbiome via probiotics, prebiotics, and fecal microbiota transplantation is gaining interest as an adjunctive approach to restoring immune homeostasis and enhancing mucosal recovery. The review also addresses the management complexities of refractory CD types I and II, which carry increased morbidity and require specialized treatment. Despite encouraging early-phase clinical trial results, none of these novel therapies have yet supplanted the GFD as standard care. Limitations include incomplete efficacy, variable patient responses, safety concerns, and challenges in long-term outcome data. Nevertheless, these advances represent significant strides toward a more comprehensive, mechanism-based approach to CD management. Future directions emphasize combination therapies, personalized medicine approaches guided by genetic and immunologic profiling, and improved biomarkers to monitor disease activity and treatment response. In summary, the evolving therapeutic landscape for coeliac disease reflects a shift from exclusive reliance on dietary gluten avoidance toward integrated strategies targeting immune pathways, intestinal barrier function, and microbial influences. These innovations hold promise to improve patient quality of life, reduce dietary burden, and ultimately achieve durable disease remission.

Keywords: biologics, gluten-free diet, immunotherapy, zonulin inhibitors

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Authors: Mohammed Alkhorayef, Abdelmoneim Sulieman, David Bradely

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Lutetium-177 (¹⁷⁷Lu) is a pivotal radionuclide in targeted radionuclide therapy (e.g., ¹⁷⁷Lu-PSMA). However, its production pathways—neutron irradiation of ¹⁷⁶Lu (direct route) or ¹⁷⁶Yb (indirect route)—generate radioactive impurities that elevate patient and staff radiation risks. Dominant impurities include long-lived ¹⁷⁷mLu (t₁/₂ = 160.4 d), high-energy gamma emitter ¹⁷⁵Yb (t₁/₂ = 4.2 d), and ¹⁷⁶Lu (t₁/₂ = 3.6 × 10¹⁰ y). Theranostic radiation dosimetry lets physicians customize treatments for each patient. Organ dose monitoring helps alter activity or treatment cycles to optimize tumor cell death while minimizing tissue harm. This study measured patient organ dosages during Lu-177 DOTATATE and PSMA theranostic procedures to assess the impurities and their relevant risks to the patient and explore the use of AI in patient dose optimisation. Nine patients received Lu-177 radionuclide treatment, generally four cycles with 6.0 to 8.1 GBq activity. The fixed activity dosage is usually 7.5 GBq. Physicians can estimate patient condition-specific dosage cycles using quantitative dosimetry. The average dosage per activity for the right kidney, left kidney, liver, bladder, and spleen was 1.76, 2.0, 0.26, 0.52, and 2.43 mSv/MBq. The left and right kidneys, liver, urinary bladder, and spleen received lower dosages than in earlier experiments. Patient organ dosages for Lu-177 DOTATATE and PSMA varied widely. More research is urgently needed to reduce radiogenic risks to healthy organs during theransotic procedures. ¹⁷⁷Lu impurities represent a challenge for clinicians and result in increased whole-body effective doses up to 0.3 mSv/MBq, mainly to non-target organs, and may result in prolonged radiotoxicity. AI can be used as an efficient tool in managing radioactive element impurities in nuclear.

Keywords: artificial intelligence, radiation risk, medical imaging, nuclear medicine

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Authors: Mohammed Alkhorayef

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Positron Emission Tomography/Computed Tomography (PET/CT) plays a crucial role in diagnosing and monitoring conditions of the lower extremities, including cancers, infections, and inflammation. Nonetheless, the radiation exposure linked to these procedures remains a key concern, particularly for patients requiring multiple scans. This study conducts an extensive dosimetric analysis of radiation doses from lower extremity PET/CT scans, evaluating both organ-specific and effective doses while exploring strategies to optimize these doses by measuring exposure levels and examining differences in protocols. Aim: This study aims to enhance clinical practices by striking a balance between diagnostic effectiveness and patient safety. Method: The study was conducted at a PET/CT research center in the Almana Hospital, Eastern Province, Dammam, Saudi Arabia. The hospital is among the country’s largest referral hospitals and oncology clinics. The dose from a combined PET/CT (Siemens Biograph mCT Flow) system, known for its precision, and the amount of radiation supplied to the patient were used to determine their exposure. The rotation time is 0.5 s, the table weight capacity is 227 kg, and the size is 78 cm. The maximum output of the CT machine is 140 kVp and 800 mA. Result: A total of 59 Lower Limb PET-CT scans were analyzed. The mean age of the patients was 54.89 years (SD = 16.1). Patient BMI was distributed across categories, with no single category exceeding 50%. The mean DLP was 1004.1 mGy.cm (SD: 287.59). The mean CTDI was 8.65 mGy (SD: 1.24). The mean radionuclide activity was 323.34 MBq (SD: 38.38). Conclusion: The results emphasize the possibility of preserving diagnostic quality in line with ALARA (As Low As Reasonably Achievable) principles.

Keywords: nuclear medicine, radiology, PET-CT, lower limb

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Authors: Akshayaa K. Aggarawal, Jithun V. Varghese, Mansoor Gazi

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Autoimmune encephalitis (AE) is a rare brain inflammation caused by the immune system. LGI1 antibody-associated AE is more common in older men and presents with memory loss, confusion, and brief seizures called facio-brachial dystonic seizures (FBDS). We report a 66-year-old man with hypertension and high cholesterol who presented with sleep-related spasms, confusion, and cognitive decline. Early MRI and EEG were normal, delaying diagnosis. LGI1 antibodies were later detected in his blood. He received corticosteroid treatment, showing slight cognitive improvement, though some symptoms like hallucinations and insomnia persisted. This case highlights the need for early testing and treatment, even when initial findings are normal.

Keywords: faciobrachial dystonic seizures, normal EEG, psychiatric symptoms, focal seizures, LGI1 antibody mediated autoimmune encephalitis

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Authors: Kelzey Nicole L. Ramos, Roy B. Tumlos, Chitho P. Feliciano

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Tumor recurrence and metastatic spread are among the adverse effects of low dose total body irradiation (TBI). However, studies revealed that increasing the dose of TBI leads to higher mortality in leukemia patients. In this study, the use of aqueous extracts was investigated to determine if these will enhance the radiation sensitivity of HL-60 leukemia cells. The water-soluble bioactives of Blumea balsamifera (sambong) leaves, and Curcuma longa (turmeric) rhizomes were extracted through decoction. The resulting sambong leaf (SLE) and turmeric extracts (TE) were tested for their antioxidant activity using the DPPH Radical Scavenging Assay and Folin-Ciocalteu Method. It was revealed that SLE has an IC50 of 0.152 mg/mL with a total phenolic content of 66.748 ± 0.753 mg GAE/g extract. TE exhibited higher IC50 (1.398 mg.mL) and lower total phenolic content (12.308 ± 0.270 mg GAE/g extract). The radiation sensitivity of HL-60 was determined by treating the cells with increasing X-ray radiation doses (0.5, 1, 2, 4, 6, and 8 Gy). The LD50 was determined to be 2.96 Gy at 4 days post-irradiation. The anti-cancer activity of SLE and TE were both evaluated, as well as the combined effects of X-ray irradiation with aqueous extracts. HL-60 cells were treated with extract concentrations of 1.5, 3, and 6 mg/mL for 24 hours. After treatment, cells were irradiated with the LD50 (2.96 Gy) and were counted at 1, 2, and 3 days post-irradiation. The findings of this study indicate that while both extracts exhibit a dose-dependent reduction of surviving cells, the aqueous extract of Curcuma longa (turmeric) rhizomes was more potent as an anti-cancer and radiosensitizing agent. These results highlight the use of natural products in the treatment of leukemia cells in vitro, warranting further investigation in vivo.

Keywords: anti-cancer, radiation sensitivity, radiosensitizing agent, water-soluble bioactives, Blumea balsamifera, Curcuma longa

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Authors: Nurul Izzati Hamzan, Hafizah Hassan, Muhammad Nursyazwan Zamre, Cefefe Stephanny Jolin, Anasufiza Habib, Siti Nurwani Ahmad Ridzuan

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Background: Congenital Disorders of Glycosylation (CDG) represent a diverse group of rare genetic disorders characterized by aberrant glycosylation, often leading to complex, multi-systemic clinical presentations. The initial biochemical screening for CDG, particularly using capillary electrophoresis (CE) to assess transferring glycoforms and Carbohydrate Deficient Transferrin (CDT) values, is crucial for identifying potential cases. However, the diagnostic implications of CE-variant patterns extend beyond confirmed CDG. Objective: This study aimed to evaluate the diagnostic accuracy of CE in identifying patients with glycosylation abnormalities and to characterize the underlying molecular heterogeneity observed in cases with variant CE patterns, encompassing both confirmed CDG and other genetic diseases. Methods: We analysed 151 patient samples, categorized into 105 normal and 46 variant-type, based on CE analysis of CDT values and sialo transferrin patterns. A subset of 21 variant samples underwent comprehensive molecular genetic testing, including a targeted CDG gene panel, whole-exome sequencing (WES), and karyotype analysis. Demographic data and quantitative CDT values were also recorded. Results: CE identified 46 samples with abnormal sialo transferrin patterns, indicative of potential glycosylation defects. Two (2) samples were confirmed to have congenital CDG. Genetic follow-up revealed notable molecular heterogeneity among these CE-variant cases. A significant portion—11 out of 21 genetically tested variant samples—were diagnosed with other rare genetic conditions unrelated to CDG. These included CHARGE syndrome, Kolen De Vries syndrome, Prader-Willi syndrome with atypical proximal breakpoint, Snijder Blok-Campeau syndrome, 3p26 microdeletion syndrome, X-linked Barth syndrome, spinocerebellar ataxia type 3 (SCA3), and Cornelia de Lange syndrome. Notably, 25 variant samples remain pending genetic analysis, and several cases that underwent whole-exome sequencing (WES) yielded inconclusive results. Conclusion: Our findings highlight the diagnostic value of capillary electrophoresis as a reliable first-line screening tool for rare diseases associated with glycosylation abnormalities. Importantly, molecular analysis of CE-variant cases revealed substantial genetic heterogeneity, with many abnormalities linked to non-CDG genetic conditions. These results emphasize that an abnormal CE pattern warrants comprehensive molecular testing to achieve an accurate diagnosis, which is essential for guiding clinical management and genetic counseling. Further research is needed to complete the genetic characterization of all CE-variant samples and to optimize diagnostic strategies for patients with atypical glycosylation profiles.

Keywords: capillary electrophoresis, glycosylation disorders, molecular heterogeneity, rare diseases

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Authors: Yung-Feng Yen

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Background: Reports of antimicrobial stewardship programs (ASP) in community hospitals are limited, with a major focus on specific agents, small settings or short periods. Here we presented the outcomes of cost control, consumption restrain and quality of care after a 3-year multidisciplinary ASP in a 415-bed community public teaching hospital. Methods: Three strategies for improving antimicrobial stewardship were implemented: education, clinical pharmacists' intervention and regular outcome announcement. The committee of infection control, composed of infection specialists, attending physicians, clinical pharmacists, nurses and medical laboratorists, was the steering panel of the program. Results: Semiannual data were presented from July 2009 to June 2012. Antibiotic cost declined from $21,464 to $12,146 in USD per 1,000 patient-days (-43.4%). Approximately $2.5 million was preserved in 3 years, and the estimated labor cost was $3,935 per month. DDDs per 1,000 patient-days were diminished from 906.7 to 717.5 (-20.9%). Significant reduction was found in the consumption of aminoglycosides, first-generation cephalosporins and aminopenicillins. However, through comprehensive auditing, increasing consumption of fourth-generation cephalosporins and fluoroquinolones was noticed. No significant difference in the quality of care (i.e., length of stay, incidence of healthcare-associated infections and mortality) was observed. Conclusion: The multidisciplinary ASP was beneficial in reducing antibiotic cost and consumption. The strategies were practical and worthy of being recommended to community healthcare settings.

Keywords: antimicrobial stewardship programs, program development, multidisciplinary, clinical pharmacists

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Authors: Muhammad A. A., Yusuf A. J., Fakurazi S.

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Background: Diabetic foot disease has become a global concern. People with diabetes have a significant percentage lifetime risk of developing foot ulcer. High prevalence rates of diabetes in many countries of the world make foot ulcers a major and increasing public-health problem. Foot ulcers are known to cause substantial morbidity, impair quality of life, endangering high treatment costs The unavailability cost effective therapeutic agents for diabetic wound healing is of great concern. The aim of this study is to perform in silico molecular docking and ADMET analysis of some bioactive compounds Identified from Moringa Oleifera against one of the important targets of wound healing protein the vascular endothelial growth factor (VEGF) towards enhancement of wound healing in diabetes.Methods: The two bioactive compounds from Moringa oleifera (Kaempferol and quercetin)earlier identified and reported in our previous study, were screened against VEGF proteins using AutoDock Vina while the ADMET analysis was performed using swissADME and admetSAR.Results: The in silico computational studies revealed that the compounds (Kaempferol and quercetin)fromMoringa oleiferacan effectively bind with high affinity and lower energy values to the VEGF which may be a target for enhancing wound healing in diabetes. ADME-Toxicity analysis revealed that, the compounds possess wound healing activity.Conclusion: The findings of this study have shown that, the plant Moringa oleifera contain effective ligands for VEGF and may therefore be considered effective in enhancing wound healing in diabetes.

Keywords: kaempferol and quercetin, diabetes, wound healing, VEGF

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Authors: Funda Bulut Arıkan, Ezgi Su Birinci

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Statement of the Problem: This study aimed to determine the distribution of immunological responses associated with food allergies and to evaluate the impact of colostrum intake and early feeding practices on the development of allergies. Methods: This cross-sectional study included 62 children aged 3–14 years, diagnosed with food allergies based on clinical history, skin prick test results, and/or specific IgE levels. Participants completed a structured questionnaire. Findings: The most frequently observed allergic symptoms were cutaneous (90%) and respiratory (43.3%) manifestations, while circulatory symptoms were the least frequent (13.3%). Ocular and gastrointestinal symptoms were seen in 36.7% of cases, and nasal and neurological symptoms in 33.3%. No significant gender differences were found in allergic responses (p>0.05). The most common allergens were nuts (50%), milk (33.3%), and eggs (26.7%), with less frequent allergens including fruits, spices, seafood, chocolate, vegetables, legumes, seeds, and wheat. A significant association was found between the age of introducing complementary foods and the development of food allergies (p=0.046). Colostrum intake was linked to a different distribution of allergic symptoms (p=0.006) and to a higher rate of exclusive breastfeeding (p=0.020). A significant relationship was also found between feeding type and the variety of allergens (p=0.034). Differences in allergy types were noted between children who received colostrum and those who did not (p=0.023). Additionally, there were significant associations between the age of complementary food introduction and allergy symptom onset (p=0.015), and between DMFT scores and allergy onset (p=0.049) and development mechanisms (p=0.001). Conclusion: These findings suggest that early complementary feeding and colostrum intake may influence the development of childhood food allergies. Colostrum intake enhances exclusive breastfeeding rates and may mitigate the severity and diversity of systemic, neurological, and circulatory symptoms by providing early immunological protection, thereby potentially reducing the risk of allergic disease development.

Keywords: allergies, colostrum, immunological response, childhood food allergies

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Authors: Malek Al Momani

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A 17-year-old previously healthy male suffered from progressive right eye proptosis and visual decline one week after a blunt head trauma. Examination revealed significant periorbital swelling, conjunctival chemosis, restricted ocular motility, and a positive relative afferent pupillary defect. Initial imaging demonstrated a subgaleal hematoma with right orbital extension, causing mass effect on the right globe. Despite emergent canthotomy and cantholysis, the patient developed progressive proptosis. Further workup revealed chronic myeloid leukemia (CML). This case underscores the importance of a multidisciplinary approach in managing orbital compartment syndrome associated with an underlying hematologic disorder.

Keywords: orbital compartment, leukemia, subgaleal hematoma, proptosis

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Authors: Gebremedhin Kide Kinfe, Birhanu Tsegaye Wores

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Background: X-ray Computed Tomography dose levels have been varying among modalities and scanning body regions due to the absence of an incessant routine follow-up. Thus, the study aimed to compute the dose index discrepancies in Ethiopia for the most recurring scan protocols (head, chest, abdomen, and pelvis). Methods: A purposive sampling method was employed to select the hospitals due to the rare existence of functional CT scanners in Ethiopia. From the selected hospitals, a total of 1,385 (249 heads, 804 chests, 132 abdomens, and 200 pelvis) were collected in terms of standard dose metric values in the period of December 2019- March 2020. Patients’ DLP was computed into a mean value using IBM SPSS Statistics 20 software. From the mean DLP, we can compute the effective dose. Results: Patients’ dose level disparity was observed in this study, though it is below the ICRP standard level for all body regions except for pelvis DLP (593.37 mGy-cm) at Black Lion. The dose level for the head and chest are computed within the recommended level at all hospitals. Effective doses for the pelvis at four hospitals (Teklehaimanot, Black Lion, ALERT, Paul’s, and Ayder hospitals) were computed as 6.45, 8.90, 5.08, 6.54, and 6.84 mSv, respectively, and the effective dose for the abdomen at Ayder Hospital was obtained to be 8.90 mSv, which is above the recommended value. Conclusion: X-ray CT scanners are somewhat properly functioning, although some sort of justification and optimization for pelvis and abdomen examinations are strongly recommended to implement as low as reasonably achievable principle.

Keywords: effective dose, ionizing radiation, dose level, Ethiopia

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Authors: Jasmyne Imani Tolber

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This paper examines how poverty affects food insecurity in Indigenous communities across North America. Indigenous households face food insecurity at much higher rates than the national average, and this research explores why that is, focusing specifically on the role poverty plays. The main aim of this study is to find how poverty contributes to food insecurity and the ways to limit access to healthy, affordable food. Using a structural inequality framework and publicly available data from sources such as the U.S. Census and the USDA, this research finds a strong connection between high poverty rates and limited access to food. It also shows how federal food programs often fall short, especially when offering culturally appropriate options. While poverty is at the center of the problem, other factors—like geographic isolation and policy barriers—make it worse. These findings suggest that improving food security in Indigenous communities will require more funding and a shift in how policies are designed and who gets to shape them. This research adds to conversations around food policy, Indigenous rights, and public health, and calls for solutions that center Indigenous leadership and food sovereignty.

Keywords: food, insecurity, indigenous, communities

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Authors: Kehinde Esther Omolabake, Christopher Busayo Olowosoke, Chioma Joy Eze, Aqsa Munir, Olusola Olatunde Dada, Grace Ayomide Oke, Nouh Mounadi, Samir Chtita, Tope Abraham Ibisanmi

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Uterine fibroids (UFs) are benign but debilitating smooth muscle tumors that affect a large proportion of women of reproductive age globally, often resulting in substantial health burdens and reduced quality of life. Despite surgical interventions remaining the predominant treatment strategy, pharmacological options remain limited due to side effects and incomplete understanding of UF pathophysiology. This study presents a comprehensive investigation of thirty-one dietary phytochemicals targeting five key protein regulators: estrogen receptor (ESR1), insulin-like growth factor 1 receptor (IGF1R), epidermal growth factor receptor (EGFR), progesterone receptor (PGR), and yes-associated protein 1 (YAP1), which have all been implicated in fibroid development and reproductive signaling. A multi-step virtual screening pipeline was employed involving molecular docking, MM/GBSA binding free energy analysis, pharmacokinetic profiling (ADMET), prediction of activity spectra (PASS), density functional theory (DFT) calculations, and 100-nanosecond molecular dynamics simulations. Compounds such as epigallocatechin gallate, genistein, quercetin, and naringin exhibited strong binding affinities (ranging from −10.1 to −14.2 kcal/mol) and favorable pharmacological properties, outperforming the reference standard drugs evaluated across binding energy, pharmacokinetic profiles, and dynamic stability at multiple protein targets. Notably, their bioactivity predictions supported anti-cancer and hormone-modulatory potential, with low electronic reactivity and stable protein-ligand interactions maintained throughout the simulation timeline. These findings provide compelling evidence for the therapeutic potential of selecting phytochemicals in modulating UF-associated molecular networks and establish a strong foundation for future experimental validation and preclinical development of anti-fibroid phytotherapeutics.

Keywords: DFT, dietary phytochemicals, molecular docking, uterine fibroids

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Authors: Abdolkarim Sheikhi, Leili Baghaie, Fatemeh Rahbarizadeh, Pooria Safarzadeh Kozani, Cobra Moradian, Mohammadreza Davidi, Narges Baharifar, Golnaz Kaboli, Mehdi Sheikhi, Yunfan Li, Mohammadamin Meghdadi, Abdulrahman M. Yaish, Aiden H. Yu, William W. Harless, Myron R. Szewczuk

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Background/Objectives: The SARS-CoV-2’s high mutations and replication rates contribute to its high infectivity and resistance to current vaccinations and treatments. The primary cause of resistance to most current treatments aligns within the coding regions for the spike S protein of SARS-CoV-2 which has mutated. As a potential novel immunotherapy, we generated a fusion protein composed of a soluble ACE2 (sACE2) linked to llama-derived anti-CD16 that targets different variants of spike proteins and enhances natural killer cells to target infected cells. Methods: Here, we generated a sACE2-AntiCD16VHH fusion protein using a Gly4Ser linker, synthesized and cloned into the pLVXEF1alpha- IRES-Puro vector, and further expressed in ExpiCHO-S cells and purified using Ni+NTA chromatography. Results: The fusion protein significantly blocked SARS-CoV-2 alpha, beta, delta, gamma, and omicron S-proteins binding and activating angiotensin-converting enzyme receptor-2 (ACE2) on ACE2-expressing RAW-Blue macrophage cells and the secretion of several key inflammatory cytokines, G-CSF, MIP-1A, and MCP-1, implicated in the cytokine release storm (CRS). The sACE2-Anti-CD16VHH fusion protein also bridged NK cells to ACE2-expressing human lung carcinoma A549 cells and significantly activated NK-dependent cytotoxicity. Conclusions: The findings show that a VHH directed against CD16 could be an excellent candidate to be linked to soluble ACE2 to generate a bi-specific molecule (sACE2-AntiCD16VHH) suitable for bridging effector cells and infected target cells to inhibit SARS-CoV-2 variant spike proteins binding to the ACE2 receptor in the RAWBlue cell line and pro-inflammatory cytokines and to activate natural killer cell cytotoxicity.

Keywords: COVID-19, SARS-CoV-2, sACE2-AntiCD16VHH, Immunotherapy

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Authors: Farhad Alipour

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Mitochondrial dysfunction is a key factor in diabetic cardiomyopathy, with AMPK playing a critical role in mitochondrial regulation. This study examined the protective effects of metformin, an AMPK activator, on high glucose–induced injury in neonatal rat cardiomyocytes. Cells were treated with high glucose (33 mM) ± metformin (2 mM), and AMPK activity was modulated using CC (inhibitor) or AICAR (activator). Cell viability, apoptosis, mitochondrial membrane potential, ROS levels, and mitochondrial morphology were assessed using standard assays. High glucose reduced AMPK activity, increased mitochondrial fragmentation and dysfunction, and triggered apoptosis. Metformin or AICAR reversed these effects, while AMPK inhibition blocked metformin’s protection. These results indicate that metformin alleviates high glucose–induced mitochondrial damage and apoptosis in cardiomyocytes via AMPK activation.

Keywords: AMPK, cardiomyocytes, mitochondria, high glucose

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Authors: Halil Asci, Arzu Ulusoy, Rumeysa Taner, Orhan Imeci, Ozlem Ozmen, Selcuk Comlekci

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Objective: This study aimed to investigate the time-dependent effects of high-voltage electric field (HV-EF) exposure on lung tissue in rats using histopathological and immunohistochemical approaches.Methods: Wistar Albino rats were randomly divided into five groups: control, 1-minute EF, 5-minute EF, 15-minute EF, and 30-minute EF exposure. Lung tissues were evaluated histologically via hematoxylin-eosin staining for hyperemia, edema, mononuclear cell infiltration, and epithelial loss. Immunohistochemical analyses were performed to assess inflammatory responses through IL-1β, RANKL, and TNF-α expression.Results: A progressive increase in histopathological damage was observed with prolonged EF exposure. The 30-minute group exhibited significantly higher scores for hyperemia, edema, mononuclear cell infiltration, and epithelial damage compared to the control and shorter exposure groups (p<0.001). Immunohistochemistry revealed markedly elevated IL-1β, RANKL, and TNF-α expression levels in the 30-minute exposure group, indicating intense inflammatory activity.Conclusion: Prolonged exposure to high-voltage electric fields induces prominent inflammatory and structural damage in lung tissue. These findings highlight the importance of defining safe exposure durations in potential clinical or environmental applications involving electric field technologies.

Keywords: electric field exposure, lung injury, histopathology, immunohistochemistry, inflammation

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Authors: Hatef Bokaei, Jaromir Hubalek, Amirmansoor Ashrafi

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Bisphenol A (BPA), a widespread endocrine-disrupting chemical found in polycarbonate plastics and epoxy resins, is increasingly linked to negative metabolic, reproductive, and developmental health effects. Yet, despite its widespread presence-measurable in over 90% of human biological samples-traditional detection methods like HPLC and mass spectrometry are too costly, technically demanding, and unavailable for large-scale or point-of-care screening. This article presents a low-cost point-of-care BPA monitoring platform by repurposing off-the-shelf commercial glucose monitors. The method relies on a four-step procedure: (1) acid-catalyzed hydrolysis of BPA-glucuronide in urine to liberate free BPA; (2) esterification of BPA with glucose at the C6 hydroxyl group to form a stable conjugate; (3) purification and separation by TLC and solvent extraction; and (4) electrochemical detection by glucose oxidase-based glucose monitors. A calibration method was developed to correlate sensor response with BPA concentrations with <5% analytical error. Proof-of-concept experiments using artificial urine showed excellent specificity, reproducibility, and conjugation stability. The platform represents an alternative application of consumer medical electronics that bridges clinical diagnostics and environmental health monitoring. Its low cost, simplicity of use, and portability make it ideally applicable to real-time exposure monitoring in both advanced and resource-limited settings, opening the door to multifunctional biosensing systems in public health infrastructure.

Keywords: biosensor, bisphenol a, point of care, glucose, esterification

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Authors: Ajinkya Rewatkar, Malla Bhaskara Rao, Arivazhagan A.

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Introduction: Lesional epilepsy is one of the major indications for resective epilepsy surgery in the current era. Objectives: This study aimed to evaluate the surgical outcomes and relevant prognostic factors in patients with low-grade epilepsy-associated neuroepithelial tumors (LEAT) and, especially, to develop a scoring system to predict postoperative seizure outcomes. Methodology: The clinical data of patients who underwent epilepsy surgery for LEAT were retrospectively studied. This is a single center observational study done between April 1st, 2000 and March 31st, 2022 at our institute. The surgical outcomes of seizure was evaluated using Recent ILAE classification. A scoring system of seizure outcomes was constructed based on the weight of the β-coefficient estimate of each predictor in the final multivariate predicting model of seizure outcomes. Results: Total of 72 patients were operated for LEAT related Drug Resistant Epilepsy. The median age at onset of seizure in this cohort was 3 years (range: 0.5 years to 74 years, median age: 5 years). there was a median follow up of 48 months from the date of surgery (mean of 47.47 ±30.183). In this study, mean number of preoperative anti-epileptic drugs was 2.86 with median of 2 with range of 1-6 drugs, and post operatively this reduced to mean of 1.86 with range of 0-4 drugs. It was found that at the end of 1st year 86.11 percent of patients were seizure free out of 72 patients. Risk probability evaluation was done and a predictive score was made. The predictive factors which were found significant were seizure duration is less than 5 years, patients do not have Acute Post Operative Seizures, if the number of pre operative Anti-epileptic drugs is less than 2 and preoperative seizure score is less than 8. Conclusions: The long-term surgical outcomes of LEAT after surgery were satisfactory. Factors predicting favourable outcome were delineated and a scoring system for predicting unfavorable seizure outcomes with different risk levels was developed, which could partly guide clinical treatments of LEAT.

Keywords: epilepsy surgery, long-term epilepsy associated tumor, neurological outcome, prognostic factor, seizure outcome

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Authors: Musefiu Yemi Adediji, Sanjeev Billa, Shabnam Siddiqui, Prabhu U. Arumugam

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Gamma-Aminobutyric Acid (GABA) and Glutamate (GLU) are key neurotransmitters involved in regulating the brain’s excitatory and inhibitory (E/I) balance. Dysregulation of these neurotransmitters contributes to neurological disorders such as epilepsy, Parkinson’s disease, dementia, and Alzheimer’s disease. Real-time monitoring of their dynamics is critical for advancing understanding and treatment strategies. Traditional methods, including high-performance liquid chromatography (HPLC) and microdialysis, offer high sensitivity but are limited by poor temporal resolution and potential tissue damage. Electrochemical biosensors based on platinum microelectrode arrays (Pt MEAs) provide minimally invasive, high-resolution alternatives capable of sub-second temporal and sub-micron spatial resolution. This study investigates electrochemical roughening (ECR) as a surface modification strategy to enhance the sensitivity of Pt MEAs for GABA and GLU detection. Three surface activation methods—alcohol activation, electrochemical cleaning (ECC), and ECR—were evaluated to optimize hydrogen peroxide (H₂O₂) detection, the reporter molecule for enzymatic biosensors. ECR was implemented using square wave voltage pulses (±1.4 V/–0.25 V) across frequencies from 0.1 to 6 kHz. Surface characterization was conducted using scanning electron microscopy (SEM), energy-dispersive spectroscopy (EDS), Raman spectroscopy, cyclic voltammetry (CV), and electrochemical impedance spectroscopy (EIS). ECR-treated electrodes demonstrated a 58% increase in voltammetric current and a 1.11% decrease in peak-to-peak potential separation, indicating enhanced electroactive surface area and improved electron transfer kinetics. EIS results confirmed a substantial reduction in charge transfer resistance. The optimized ECR process achieved peak in vitro sensitivities of 6810 ± 103 nA µM⁻¹ cm⁻² for H₂O₂, 45 ± 4.38 nA µM⁻¹ cm⁻² for GABA, and 1510 ± 47 nA µM⁻¹ cm⁻² for GLU, with low detection limits of 0.00160 µM for GABA and 0.0127 µM for GLU. These results highlight the potential of ECR-enhanced Pt MEAs for high-sensitivity neurochemical monitoring.

Keywords: electrochemical biosensor, E/I balance, GABA, glutamate, hydrogen peroxide, platinum microelectrodes, surface roughening

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