https://hdl.handle.net/2072/478782 Wed, 08 Apr 2026 02:22:22 GMT 2026-04-08T02:22:22Z https://hdl.handle.net/2445/223805 Inhibition of PCSK9 with polypurine reverse hoogsteen hairpins: A novel gene therapy approach López-Aguilar, Ester; Pacheco-Velázquez, Silvia Cecilia; Busquets i Viñas, Ma. Antonia; Hay, J.oshua; Mueller, P.aul A.; Fazio, Sergio; Ciudad i Gómez, Carlos Julián; Noé Mata, Verónica; Pamir, Nathalie PCSK9 is a therapeutic target for hypercholesterolemia. Though different strategies to inhibit PCSK9, such as monoclonal antibodies, small molecules, or nucleic acid drugs are available, the need for safer and inexpensive interventions remains. We developed a time-, cost-, and resource- efficient silencing system using Polypurine Reverse Hoogsteen (PPRH) hairpins to target PCSK9. To achieve PCSK9 silencing, we designed two PPRHs targeting PCSK9 at exon 9 (HpE9) and exon 12 (HpE12). The binding capabilities of PPRHs were measured by EMSA: Kd values were 7.86 x 10-8 M and 7.58 x 10-7 M for HpE9 and HpE12, respectively. PPRHs were complexed with the cationic polymer jetPEI forming particles of 167 nm as characterized by Dynamic Light Scattering. PCSK9 gene and protein expression was evaluated upon transfections of HepG2 cells with HpE9 and HpE12. PPRHs effectively reduced PCSK9 mRNA levels (63 % and 74 % for HpE9 and HpE12, respectively) and protein (by 76 % and 87 %) at 24 h. Human PCSK9 overexpressing mice receiving a single injection of HpE12 decreased plasma PCSK9 levels by 50 % by day three post injection and levels returned to baseline by day fifteen. Plasma cholesterol levels were reduced by 47 % by day three. Mice receiving the PPRHs did not exhibit changes in body weight, liver enzymes or pro-inflammatory markers when compared to mice injected with jetPEI alone. Therefore, the PPRH technology emerges as an innovative nucleic acid based therapeutic approach that is effective, cost-efficient and easy to develop, for the inhibition of PCSK9. Wed, 22 Oct 2025 07:20:27 GMT https://hdl.handle.net/2445/223805 2025-10-22T07:20:27Z https://hdl.handle.net/2445/222310 Assessment of Flurbiprofen Suspension and Composite GelPre- and Post Skin Perforation: Effectiveness in ManagingInflammatory Responses in Ear Tags and Periocular Piercings. El Bejjaji, Sheimah; Ramos Yacasi, Gladys Rosario; Domínguez Villegas, Valeri; Chaves Moreira Dos Santos, Délia; Braza Reyes, Antonio J.; Sosa Díaz, Lilian Elisa; Rodríguez Lagunas, María José; Calpena Campmany, Ana Cristina; Zelaya, Mireia; Parra Coca, Alexander Background: Controlled skin perforations, such as ear tags, piercings, and microdermal implants, induce inflammation and stress in individuals undergoing these procedures. This localized trauma requires care to optimize healing, reduce inflammation, and prevent infections. (2) Methods: Two formulations were developed: an FB-suspension and an FB-gel. Their in vivo efficacy was evaluated, along with drug retention in porcine and human skin after 30 min of administration, chemical stability at different temperatures, cytotoxicity, histological changes induced via transdermal application, and irritative potential, assessed using the HET-CAM assay. (3) Results: Both formulations reduced inflammation when applied 30 min before perforation compared to the positive control. The FB-suspension demonstrated no cytotoxicity and exhibited greater efficacy than the free flurbiprofen solution, highlighting the advantages of using nanoparticle-mediated drug delivery. Moreover, the FB-gel maintained chemical stability for up to 3 months across a temperature range of 4 to 40 °C. Histologically, no significant changes in skin composition were observed. (4) Conclusions: The FB-suspension is viable for both pre- and post-perforation application, as it is a sterile formulation. In contrast, the FB-gel is a convenient and easy application, making it a practical alternative for use in both clinical and veterinary settings. Keywords: flurbiprofen; suspension; hydrogel; polyethylene glycol 3350; human skin; porcine skin; NSAID; transdermal drug delivery; controlled skin perforation; inflammation management Thu, 17 Jul 2025 07:23:26 GMT https://hdl.handle.net/2445/222310 2025-07-17T07:23:26Z https://hdl.handle.net/2445/221705 Developing a global education hub for animal-free innovation Janssens, Monique R. E.; Salvatori, Daniela; Hogervorst, Janneke; Nonis, Cristheena; Bailey, Jarrod; Bajramovic, Jeffrey; Burgers, Anne; Caloni, Francesca; van Deel, Elza D.; van den Eijnden-van Raaij, Janny; Amirabadi, Hossein E.; Filipova, Dilyana; Gastaldello, Annalisa; Gibbs, Susan; Goversen, Birgit; Green, Nicole; van Hengel, Jolanda; Kienhuis, Anne; van de Kolk, Sjoukje; Paggi, Carlo A.; Penning, Louis C.; Pistollato, Francesca; Riegger, Silke; Ritskes-Hoitinga, Merel; Vinardell Martínez-Hidalgo, Ma. Pilar In recent decades, the life sciences have witnessed remarkable advancements, leading to breakthroughs in medicine, agriculture, and environmental science. Although experiments on animals have been used on the way to making these advancements, the scientific community and society are increasingly questioning the scientific validity and ethics of using animals in research, testing, teaching, and training. Systematic reviews have shown that the translatability of results from animal studies to humans is often poor (Leenaars et al., 2019), and the use of animals in experiments is often termed “a black box” because the mechanisms at work are unclear. In addition, experiments using genetically homog-enous strains of animals do not reflect the interindividual differ-ences among patients. Diseases are often induced in experimental animals in an artificial way, for example by genetic modification or chemical insult, which means that the human etiology of the diseases is not accurately represented, hindering opportunities to identify avenues for prevention and treatment. Mon, 23 Jun 2025 07:33:11 GMT https://hdl.handle.net/2445/221705 2025-06-23T07:33:11Z https://hdl.handle.net/2445/205847 Mendelian Randomization Analysis of the Relationship Between Native American Ancestry and Gallbladder Cancer Risk Zollner, Linda; Boekstegers, Felix; Barahona Ponce, Carol; Scherer, Dominique; Marcelain, Katherine; Gárate-Calderón, Valentina; Waldenberger, Melanie; Morales, Erik; Rojas, Armando; Muñoz, Cesar; Müller, Bettina; Retamales, Javier; de Toro, Gonzalo; Kortmann, Allan Vera; Barajas, Olga; Rivera, María Teresa; Cortés, Analía; Loader, Denisse; Saavedra, Javiera; Gutiérrez, Lorena; Ortega, Alejandro; Bertrán, Maria Enriqueta; Bartolotti, Leonardo; Gabler, Fernando; Campos, Mónica; Alvarado, Juan; Moisán, Fabrizio; Spencer, Loreto; Nervi, Bruno; Carvajal-Hausdorf, Daniel E.; Losada, Héctor; Almau, Mauricio; Fernández, Plinio; Olloquequi, Jordi; Carter, Alice R.; Miquel Poblete, Juan Francisco; Bustos, Bernabe I.; Fuentes-Guajardo, Macarena; Gonzalez-Jose, Rolando; Bortolini, M. C.; Acuña-Alonzo, Víctor; Gallo, Carla; Ruiz Linares, Andres; Rothhammer, Francisco; Lorenzo Bermejo, Justo Background A strong association between the proportion of Native American ancestry and the risk of gallbladder cancer (GBC) has been reported in observational studies. Chileans show the highest incidence of GBC worldwide, and the Mapuche are the largest Native American people in Chile. We set out to investigate the causal association between Native American Mapuche ancestry and GBC risk, and the possible mediating effects of gallstone disease and body mass index (BMI) on this association. Methods Markers of Mapuche ancestry were selected based on the informativeness for assignment measure and then used as instrumental variables in two-sample mendelian randomization (MR) analyses and complementary sensitivity analyses. Result We found evidence of a causal effect of Mapuche ancestry on GBC risk (inverse variance-weighted (IVW) risk increase of 0.8% for every 1% increase in Mapuche ancestry proportion, 95% CI 0.4% to 1.2%, p = 6.6×10-5). Mapuche ancestry was also causally linked to gallstone disease (IVW risk increase of 3.6% per 1% increase in Mapuche proportion, 95% CI 3.1% to 4.0%, p = 1.0×10-59), suggesting a mediating effect of gallstones in the relationship between Mapuche ancestry and GBC. In contrast, the proportion of Mapuche ancestry showed a negative causal effect on BMI (IVW estimate -0.006 kg/m2 per 1% increase in Mapuche proportion, 95% CI -0.009 to -0.003, p = 4.4×10-5). Conclusions The results presented here may have significant implications for GBC prevention and are important for future admixture mapping studies. Given that the association between Mapuche ancestry and GBC risk previously noted in observational studies appears to be causal, primary and secondary prevention strategies that take into account the individual proportion of Mapuche ancestry could be particularly efficient. Wed, 17 Jan 2024 12:47:52 GMT https://hdl.handle.net/2445/205847 2024-01-17T12:47:52Z