Background/Objectives: Boron neutron capture therapy (BNCT) is a promising approach for selectively targeting and destroying malignant cells using ¹⁰B isotopes. A significant challenge in BNCT lies in the development of efficient boron delivery systems that ensure adequate boron accumulation within tumor cells. This study aims to synthesize, characterize, and evaluate COSAN-functionalized nanoparticles (NP@I-COSAN) as a potential boron carrier for BNCT. Methods: Hybrid nanoparticles were synthesized by conjugating monoiodinated cobaltabisdicarbollides (I-COSAN) to commercially available acrylic polymer-based nanoparticles. Functionalization and cellular uptake were confirmed through FTIR, TGA, UV-Vis spectroscopy, and TEM/EDX analyses. Biocompatibility was evaluated by assessing cytotoxicity in HeLa cells and C. elegans as an in vivo model. Intracellular boron uptake was quantified using ICP-MS, with results compared to those obtained with 4-borono-L-phenylalanine conjugated to fructose (f-BPA). An in vitro BNCT proof-of-concept assay was also performed to evaluate therapeutic efficacy. Results: NP@I-COSAN demonstrated low cytotoxicity and efficient internalization in cells. ICP-MS analysis revealed stable boron retention, comparable to traditional boron agents. The BNCT assay further showed that NP@I-COSAN was effective in inducing tumor cell apoptosis, even at lower boron concentrations than conventional treatments. Conclusions: These results underscore the potential of NP@I-COSAN as an effective boron delivery system for BNCT, offering a promising strategy to enhance boron accumulation within tumor cells and improve treatment efficacy.

Parkinson's disease (PD) is among the most prevalent neurodegenerative disorders and is diagnosed based on motor symptoms that emerge after significant dopaminergic neuron loss. Non-motor symptoms, such as constipation, are the most common and often appear earlier, indicating the onset of the disease. Many animal models are used to study neurodegenerative diseases, yet most focus on the later stages. Because it is difficult to determine the exact start of neurodegeneration, no single model fully represents the entire disease progression. Mammalian models are especially time-consuming and expensive due to their long-life cycles. In contrast, Caenorhabditis elegans (C. elegans), a transparent 1 mm nematode with a short life and reproductive cycle (~3 weeks and 3 days), enables faster, cost-effective experiments thanks to its rapid generation turnover. The defecation motor program in C. elegans consists of rhythmic cycles regulated by a complex genetic network in which Rho/rac GTPases participate, and RAC1 (CED-10 in C. elegans) is among them. We have shown that RAC1/CED-10 mutants exhibit compromised GABAergic morphology and function, which results in disrupted defecation cycles. Here, we present the experimental protocol for measuring constipation in RAC1/CED-10 C. elegans at early stages. In addition to serving as a predictive tool to investigate early PD symptoms prior to neurodegeneration, this protocol could also be employed as an early endpoint to assess the efficacy of pharmacological intervention. Because these changes are not disease-specific, we believe this model provides a valuable tool for investigating alterations that arise early in other illnesses.

Parkinson's disease (PD) is among the most prevalent neurodegenerative disorders and is diagnosed based on motor symptoms that emerge after significant dopaminergic neuron loss. Non-motor symptoms, such as constipation, are the most common and often appear earlier, indicating the onset of the disease. Many animal models are used to study neurodegenerative diseases, yet most focus on the later stages. Because it is difficult to determine the exact start of neurodegeneration, no single model fully represents the entire disease progression. Mammalian models are especially time-consuming and expensive due to their long-life cycles. In contrast, Caenorhabditis elegans (C. elegans), a transparent 1 mm nematode with a short life and reproductive cycle (~3 weeks and 3 days), enables faster, cost-effective experiments thanks to its rapid generation turnover. The defecation motor program in C. elegans consists of rhythmic cycles regulated by a complex genetic network in which Rho/rac GTPases participate, and RAC1 (CED-10 in C. elegans) is among them. We have shown that RAC1/CED-10 mutants exhibit compromised GABAergic morphology and function, which results in disrupted defecation cycles. Here, we present the experimental protocol for measuring constipation in RAC1/CED-10 C. elegans at early stages. In addition to serving as a predictive tool to investigate early PD symptoms prior to neurodegeneration, this protocol could also be employed as an early endpoint to assess the efficacy of pharmacological intervention. Because these changes are not disease-specific, we believe this model provides a valuable tool for investigating alterations that arise early in other illnesses.