Search

Jessica Perez, Javier Cifuentes, Monica Cuellar, Alejandra Suarez-Arnedo, Juan C Cruz, Carolina Muñoz-Camargo GINIB Research Group, Department of Biomedical Engineering, Universidad de los Andes, Bogotá, ColombiaCorrespondence: Carolina Muñoz-Camargo; Juan C Cruz Tel +57-1 3394949 Ext. 1789, 1853Email c.munoz2016@uniandes.edu.co; jc.cruz@uniandes.edu.coIntroduction: Controlled delivery of therapeutic molecules in a localized manner has become an area of interest due to its potential to reduce drug exposure to healthy tissues and consequently to minimize undesirable side effects. We have recently introduced novel cell-penetrating vehicles by immobilizing the antimicrobial peptide Buforin II (BUF-II) on magnetite nanoparticles (MPNPs). Despite the potent translocating abilities of such nanobioconjugates, they failed to preserve the antimicrobial activity of native BUF-II. In this work, we explored immobilization on MNPs with the aid of polymer surface spacers, which has been considered as an attractive alternative for the highly efficient conjugation of various biomolecules.Methods: Here, we immobilized BUF-II on polyetheramine-modified magnetite nanoparticles to preserve its structural integrity. As a result, for the obtained nanobioconjugates the lost antimicrobial activity against gram-positive and gram-negative bacteria was only 50% with respect to the native BUF-II. The nanobioconjugates were also characterized via FTIR, DLS, TEM, and TGA. Delivery on THP-1, HaCaT, HFF, and Escherichia coli cells was conducted to confirm capability for cell membrane translocation.Results: Colocalization with Lysotracker showed an endosomal escape efficiency of about 73∓12% in THP-1 cells. Avoidance of endocytic pathways of internalization was qualitatively confirmed by a delivery assay at low temperature. Nuclear penetration of the nanobioconjugates was corroborated via confocal microscopy and showed high biocompatibility as demonstrated by hemolysis levels below 5% and acute cytotoxicity of around 15%.Conclusion: The obtained nanobioconjugates were capable of translocating the cell membrane and nuclei of different normal and cancerous cell lines without significantly decreasing viability. This makes the vehicle addressable for a number of applications ranging from antimicrobial topical treatments to the delivery of nucleotides and therapeutic molecules with difficulties to bypass cell membranes.Keywords: Buforin-II conjugates, drug delivery, translocation, cell-penetrating peptides, nanomaterials

Monica Cuellar,1 Javier Cifuentes,1 Jessica Perez,1 Alejandra Suarez-Arnedo,1 Julian A Serna,1 Helena Groot,2 Carolina Muñoz-Camargo,1 Juan C Cruz1 1Department of Biomedical Engineering, Universidad de los Andes, Bogotá, Colombia; 2Human Genetics Laboratory, Department of Biological Sciences, Universidad de los Andes, Bogotá, Colombia Introduction: One of the major challenges of modern pharmacology is the development of systems for the delivery of therapeutic molecules in a controlled and localized manner. One strategy is to use nanostructured supports, which are well suited to carry a large number of molecules on a per mass basis. A major challenge for these supports is, however, their limited ability to bypass the cell membrane. Recent studies propose that to overcome this issue, potent translocating cell-penetrating peptides (CPPs) can be conjugated to their surfaces. Methods: Here, we conjugated the antimicrobial CPP buforin II (BUF2) to the surface of magnetite nanoparticles to enhance their cell penetration. Conjugates were characterized via Fourier transform infrared spectroscopy, dynamic light scattering, and thermogravimetric analysis, and their biocompatibility was corroborated. The conjugates were delivered in both bacterial and mammalian cells demonstrating the intracellular inclusion in THP-1 cells for the first time. Results: Despite the promising outcome, our studies showed that the obtained conjugates failed to maintain the native antimicrobial activity of BUF2. We hypothesize that to overcome this issue, a flexible linker can be inserted prior to conjugation. Conclusion: Our study highlights the potential of BUF2-magnetite conjugates as cell-penetrating vehicles for the targeted delivery of pharmacological agents. This provides support for the idea of a promising combined drug delivery and antimicrobial peptide therapy. Keywords: BUF2-magnetite, antimicrobial activity, cell-penetrating peptides, nanomaterials, drug delivery

Des recherches ont été effectuées concernant la zooépibiose de l’écrevisse signal (Pacifastacus leniusculus, D.), provenant d’un centre d’astaciculture espagnol, au cours de la période 1997-1998. Les organismes épibiotiques dominants trouvés sur l’exosquelette appartiennent aux groupes zoologiques d’Invertébrés suivants : Mastigophora (Bodo), Cilliophora (Stentor, Epistylis, Zoothamnium, Acineta), Turbelaria (Temnocephala), Rotifera (Brachionus), Annelida (Branchiobdella), Acarina (Hygrobates) et Ostracode (Eucypris ou Enthocytera), on souligne le nombre élevé de Branchiobdellides de l’espèce Branchiobdella instabilia M., dont les quantités minimum et maximum, en mâles et femelles, oscillent de 12 à 139 et de 7 à 130, respectivement. Les hyperbranchiobdelloses provoquent des automutilations, des décollements des pinces et autres appendices, des mues anormales, des lésions cuticulaires, de l’hémocytopénie et des changements éthologiques. Les colonisations maximales d’Ostracodes de l’espèce Eucypris virens, J.O., Enthocytera heterodonta, R., dans les poches d’incubation des femelles donnent lieu à une coloration rougeâtre des oeufs et à leur perte postérieure. Compte tenu de leur nombre limité et de la période d’épibiose, les genres des sous-types Mastigophora, Ciliophora, Turbelaria, Rotifera et Arachnida ne représentent aucun danger pour les écrevisses. En ce qui concerne la zooépibiose associée aux appendices et aux régions du corps des écrevisses du Pacifique, les chélipèdes ou pinces constituent le substrat où se place de préférence le genre Branchiobdella ; le céphalothorax sert de support à tous les groupes taxonomiques d’épibiontes identifiés.

Background: The Rio Grande Valley (RGV) and Laredo regions located along the Texas-Mexico border consist of seven counties with a population of approximately 1.5 million people and a high uninsured rate (33.5%). Cervical cancer mortality in these border counties is approximately 30% higher than the rest of Texas. The RGV and Laredo areas were studied to better understand the state of access to cervical cancer prevention services along the Texas-Mexico border.Methods: Data on the population served and the services provided were analyzed to determine the gap between cervical cancer screenings recommended versus those received. Through interviews, we gathered the perspectives of 16 local stakeholders regarding cervical cancer screening for underserved individuals in the region.Findings: It is estimated that 69,139 uninsured women aged 21-64 years in the RGV/Laredo per year are recommended to undergo cervical cancer screening with Papanicolaou (Pap) and/or human papillomavirus (HPV) testing, but only 8941 (12.9%) Pap tests are being performed by the Federally Qualified Health Center (FQHC) serving uninsured women in these regions. Systemic barriers identified include insufficient provider clinical capacity, the high cost of healthcare, and uncertainty about government funding sources. Patient barriers identified include inadequate knowledge on navigating the local healthcare system, low health literacy, lack of money and childcare, an inability to miss work, limited transportation, and fear of deportation.Conclusion: Decreasing the disparity between cervical cancer screening services provided and those recommended requires addressing the barriers, identified by local experts, which prevent uninsured women from accessing care. These challenges are being addressed through ongoing programs and collaborations. [ABSTRACT FROM AUTHOR]