Biotechnology Research Center

There are typically three treatment options for superficial abnormal mucosal tissue, such as Barrett's esophagus: medication, endoscopic procedures, and surgery. The choice of treatment depends on the severity of the condition. Among these options, endoscopic ablation techniques have gained recognition as the most effective. However, achieving the optimal depth of therapy to ensure complete eradication of lesions remains a significant challenge. Insufficient depth can lead to recurrence or allow lesions to remain hidden beneath healthy tissue, while excessive depth increases the risk of complications such as bleeding, perforation, or organ narrowing. In this study, we introduced a design for an endocap that efficiently confines laser energy within a recessed channel. This design allows for precise angular control, enabling welldefined radial coagulation of the esophageal mucosal tissue. To implement this design, we used a specially crafted rigid insertion tube for accurate photothermal coagulation of the mucosal tissue. The endocap is equipped with a GRIN lens, a stepper motor, and gold-coated right-angle prism mirrors. We conducted tests at various laser scanning speeds and rotation angles while sliding the cap over the surface of a sheep esophagus model. Our findings suggest that this endocap design has the potential to yield more effective and safer treatment options for superficial esophageal abnormalities.

Abstract Abstract Doxorubicin (DOX) is a highly effective chemotherapeutic agent limited by dose-dependent cardiotoxicity. A key pathogenic event involves mitochondrial accumulation in cardiomyocytes, however, the resulting metabolic adaptations remain poorly understood. Therefore, this study characterizes DOX-induced alterations in cardiac metabolism and highlights their potential as novel therapeutic targets. Methods Female BALB/c mice received DOX over two weeks (12 mg/kg) and were sacrificed at early (3 days) or late (6 weeks) timepoints. Cardiac metabolism was assessed by metabolomics, 13C₆-glucose flux analysis, proteomics and metabolic enzymes activity in whole hearts and isolated mitochondria. Oxygen consumption was measurd in vitro (Seahorse) and ex vivo (Oroboros). The contribution to DOX-induced metabolic rewiring of PI3Kγ, that has been previously showed to be activated by DOX, was studied in mice expressing a kinase-inactive enzyme (PI3Kγ kinase-dead; KD). To unravel the role of PDK4-PDH axis, PDK4 overexpression was induced in mice using AAV9 viral vector, followed by DOX treatment. Results Following acute DOX administration, hearts exhibited an unexpected hypermetabolic response characterized by increased mitochondrial respiration and enhanced substrate flexibility, particularly for glucose, as confirmed by 13C₆-glucose tracing. Untargeted metabolomics revealed pyruvate depletion and acetyl-CoA accumulation, accompanied by altered protein acetylation, as evidenced by proteomics for lysine acetylation, and buildup of TCA intermediates. These changes, together with increased glutamine utilization reminiscent of cancer cell metabolism, marked an acute adaptive response. However, the persistence of this hypermetabolic state led to maladaptation, resulting in mitochondrial overload, energetic uncoupling, oxidative stress, and ultimately cardiac dysfunction. Notably, inhibition of the PI3Kγ–PDK4–PDH axis, either through PI3Kγ blockade or PDK4 overexpression, fully prevented chronic metabolic alterations, identifying PI3Kγ as a key regulator of substrate selection and metabolic remodeling during DOX treatment. Metabolic rewiring also involved cardiac fibroblasts, which showed reduced glycolysis, altered lipid handling, and increased glutamine metabolism, promoting cytosolic acetyl-CoA production via ACLY in the early phase. This shift may drive sustained fibroblast activation in the long-term, through post-translational and epigenetic mechanisms, contributing to cardiotoxicity via metabolic crosstalk with cardiomyocytes and induction of senescence. Conclusions Overall, this work demonstrates that DOX induces an acute increase in metabolic activity, with a reprogramming of different cardiac cells metabolism. While initially adaptive, this response becomes maladaptive over time, ultimately leading to chronic cardiac dysfunction.

Thymoquinone (TQ), a bioactive compound derived from Nigella sativa herbal plant, exhibits antioxidative, anti-inflammatory, and antimicrobial effects against various bacteria, viruses, fungi, protozoa, and flatworms. A systematic search was conducted across Embase, PubMed, Scopus, and Web of Science until January 2025. All preclinical studies discussing the bactericidal, virucidal, fungicidal, and parasiticidal effects of nano-based formulations of N. sativa/TQ were included. Overall, 148 studies—comprising in vitro, in vivo, ex vivo, and in silico research—were identified involving 185 unique N. sativa/TQ-loaded nanocarriers with antimicrobial properties. The predominant nanoencapsulation methods included inorganic nanoparticles (27%), nanocomposites (23.2%), and nanofibers (13.5%). The essential oil (27.6%) and aqueous extract (20.5%) of the seeds, along with TQ (22.2%), were the most common natural cargos. 149 nanoformulations showed effectiveness against bacteria, 42 against fungi, 8 against viruses, and 7 against parasites. Additionally, 29 nanoformulations showed antibiofilm properties, and 41 showed efficacy against multidrug-resistant pathogens. Only 18 nanoformulations were not effective against at least one tested pathogen. 91.9% of the tested nanoformulations against Staphylococcus aureus showed effectiveness, while this percentage for Escherichia coli and Candida albicans was 88.5% and 96.9%, respectively. These three were the most widely examined pathogens in preclinical studies. The proposed clinical and practical applications were most prominent in wound healing (42 nanoformulations), followed by food preservation (33 nanoformulations) and water purification (9 nanoformulations). Integrating nano-based drug delivery systems with the antimicrobial benefits of TQ can address TQ’s limitations, including instability and low oral bioavailability. This review identifies knowledge gaps regarding TQ nanomedicines and emphasizes the need for regulatory frameworks for safety and clinical applicability, thereby facilitating their progression into human clinical trials.

The efficacy of breast cancer chemotherapies is frequently limited by multidrug resistance (MDR), partly through efflux by ABC transporters, including ABCG2. This study evaluated whether phaeosphaeride A (PPA), a fungal metabolite isolated from a Paraphoma sp. endophyte of Ferula xylorhachis, can modulate ABCG2-mediated resistance to mitoxantrone (MTX). The endophyte was cultured and extracted with ethyl acetate, and the extract was purified by column chromatography and HPLC to yield PPA, whose structure was confirmed by NMR and MS analyses. Cytotoxicity of MTX, PPA, and their combination was assessed in MCF-7- and ABCG2-overexpressing MCF-7/MX cells. MTX showed marked differential cytotoxicity (IC50 = 1.6 µM, 95% CI: 1.4 - 1.9 in MCF-7 vs. > 25 µM in MCF-7/MX; p < 0.0001), whereas PPA exhibited comparable activity in both lines (23.2 µM, 95% CI: 18.0 - 30.1 vs. 36.1 µM, 95% CI: 28.7 - 46.0; p = 0.01). Co-treatment with PPA IC50 significantly reduced MTX IC50 to 0.4 µM (95% CI: 0.3 - 0.6) in MCF-7 and 1.9 µM (95% CI: 1.2 - 2.7) in MCF-7/MX, restoring MTX sensitivity in resistant cells to near MCF-7 levels. Flow cytometry showed that PPA (IC50) increased intracellular MTX accumulation with stronger effects in MCF-7/MX cells (p < 0.0001) than in MCF-7 (p < 0.05). In combination with MTX, PPA (IC50) increased Sub-G1 apoptotic populations in both lines.These findings demonstrate that PPA is unlikely to be a substrate of ABCG2 but functionally inhibits ABCG2-mediated efflux, contributing to the restoration of MTX sensitivity, although there may be additional mechanisms involved. PPA could be a promising MDR-reversal agent in ABCG2-driven chemotherapy resistance.

Oxidative stress-induced cellular damage is a primary contributor to pathological conditions such as accelerated skin aging. Consequently, there is a strong demand for efficacious antioxidants. Herein, we investigate the cytoprotective properties of Crocodylus siamensis white blood cell extracts and peptides derived from them against H2O2-induced oxidative damage in HaCaT keratinocytes. The extracts show no cytotoxicity, restore cell viability by 3-fold relative to oxidatively stressed cells, and reduce apoptosis. Notably, two enriched peptidesNV10 and RI10demonstrated superior cytoprotective efficacy under oxidative stress. These peptides scavenge intracellular ROS (∼60% reduction), modulate Keap1-Nrf2 pathway components at mRNA levels (downregulating Keap1, upregulating Nrf2), and suppress apoptosis via regulation of caspase and Bcl-2 mRNA expression. Molecular docking predictions suggest that the peptides bind to Keap1, a negative regulator of Nrf2, thereby providing a potential mechanism for Nrf2 pathway activation. These findings suggest that the identified peptides are potential candidates for therapeutic development in skin protection applications.