Tailoring Magnetite-Nanoparticle-Based Nanocarriers for Gene Delivery: Exploiting CRISPRa Potential in Reducing Conditions.

Gene delivery has emerged as a promising alternative to conventional treatment approaches, allowing for the manipulation of gene expression through gene insertion, deletion, or alteration. However, the susceptibility of gene delivery components to degradation and challenges associated with cell penetration necessitate the use of delivery vehicles for effective functional gene delivery. Nanostructured vehicles, such as iron oxide nanoparticles (IONs) including magnetite nanoparticles (MNPs), have demonstrated significant potential for gene delivery applications due to their chemical versatility, biocompatibility, and strong magnetization. In this study, we developed an ION-based delivery vehicle capable of releasing linearized nucleic acids (tDNA) under reducing conditions in various cell cultures. As a proof of concept, we immobilized a CRISPR activation (CRISPRa) sequence to overexpress the pink1 gene on MNPs functionalized with polyethylene glycol (PEG), 3-[(2-aminoethyl)dithio]propionic acid (AEDP), and a translocating protein (OmpA). The nucleic sequence (tDNA) was modified to include a terminal thiol group and was conjugated to AEDP's terminal thiol via a disulfide exchange reaction. Leveraging the natural sensitivity of the disulfide bridge, the cargo was released under reducing conditions. Physicochemical characterizations, including thermogravimetric analysis (TGA) and Fourier-transform infrared (FTIR) spectroscopy, confirmed the correct synthesis and functionalization of the MNP-based delivery carriers. The developed nanocarriers exhibited remarkable biocompatibility, as demonstrated by the hemocompatibility, platelet aggregation, and cytocompatibility assays using primary human astrocytes, rodent astrocytes, and human fibroblast cells. Furthermore, the nanocarriers enabled efficient cargo penetration, uptake, and endosomal escape, with minimal nucleofection. A preliminary functionality test using RT-qPCR revealed that the vehicle facilitated the timely release of CRISPRa vectors, resulting in a remarkable 130-fold overexpression of pink1. We demonstrate the potential of the developed ION-based nanocarrier as a versatile and promising gene delivery vehicle with potential applications in gene therapy. The developed nanocarrier is capable of delivering any nucleic sequence (up to 8.2 kb) once it is thiolated using the methodology explained in this study. To our knowledge, this represents the first MNP-based nanocarrier capable of delivering nucleic sequences under specific reducing conditions while preserving functionality.

Understanding the Potential of Genome Editing in Parkinson's Disease.

CRISPR is a simple and cost-efficient gene-editing technique that has become increasingly popular over the last decades. Various CRISPR/Cas-based applications have been developed to introduce changes in the genome and alter gene expression in diverse systems and tissues. These novel gene-editing techniques are particularly promising for investigating and treating neurodegenerative diseases, including Parkinson's disease, for which we currently lack efficient disease-modifying treatment options. Gene therapy could thus provide treatment alternatives, revolutionizing our ability to treat this disease. Here, we review our current knowledge on the genetic basis of Parkinson's disease to highlight the main biological pathways that become disrupted in Parkinson's disease and their potential as gene therapy targets. Next, we perform a comprehensive review of novel delivery vehicles available for gene-editing applications, critical for their successful application in both innovative research and potential therapies. Finally, we review the latest developments in CRISPR-based applications and gene therapies to understand and treat Parkinson's disease. We carefully examine their advantages and shortcomings for diverse gene-editing applications in the brain, highlighting promising avenues for future research.

Multidisciplinariedad, interdisciplinariedad y transdisciplinariedad en la formación para la investigación en ingeniería *

En el presente texto, se dan a conocer aspectos relevantes de la formación en investigación en la Facultad de Ingeniería de la Corporación Universitaria Americana. En primer lugar, se estudian los conceptos de Multidisciplinariedad, Interdisciplinariedad y Transdisciplinariedad desde su relación con la formación. En segundo lugar, se identifica el nivel de influencia de las percepciones de estudiantes en los procesos de formación en Investigación a la luz de la Estrategia de Formación por Proyectos mediante un análisis exploratorio de los resultados obtenidos de una encuesta de evaluación de una actividad transdisciplinar denominada "foro de lecciones aprendidas". Como conclusión de los resultados de la investigación, se identifica la independencia entre semestre y el concepto que los estudiantes tienen acerca de su formación por competencias en relación al foro y se proponen estrategias de intervención en este aspecto.

Introduction. Nowadays, engineering schools are faced with complex needs in realities that are often alien to them and transcend disciplinary limits. The efforts to understand this result in the implementation of conceptualization spaces oriented towards the articulation of processes coherent with the institutional work and the policies of accreditation in high quality. Objective. To analyze relevant aspects of research training in the School of Engineering of the American University Corporation considering the challenges imposed by the disciplinary, multidisciplinary interdisciplinary, and / or transdisciplinary tensions of sciences. Materials and methods. Disciplinarity, Multidisciplinarity, Interdisciplinarity, and Transdisciplinarity concepts are studied from their relationship with training. Subsequently, perceptions of students in research training processes are identified through an exploratory analysis of results obtained from an evaluation survey of a transdisciplinary activity called "lessons learned forum," the analysis was carried out using a chi-square test with a 95% reliability for a sample of 200 students of the Engineering School for the year 2016. Results. Taking into account the rejection of independence for 24 2 36,4 , in which 24 represent the degrees of freedom with a 5% error, the initial hypothesis is retained. Conclusion: There is an independence between the semester and students' concept about their training by competences in relation to the forum, which entails the need to propose intervention strategies for the transgression of what is disciplinary and to promote complex thinking in students.

Introdução. Atualmente, as faculdades de engenharia se encontram ante necessidades complexas em realidades que muitas vezes são alheias a elas e transcendem os limites do disciplinar. Os esforços por compreender este, derivam na implementação de espaços de conceptualização orientados para a articulação de processos coerentes com o trabalho institucional e as políticas de acreditação em alta qualidade. Objetivo. Analisar aspectos relevantes da formação em investigação na Faculdade de Engenharia da Corporação Universitária Americana considerando os desafios que impõem as tensões entre o disciplinar, multidisciplinar, interdisciplinar e/ou transdisciplinar das ciências. Materiais e métodos. Se estudam os conceitos de Disciplinariedade, Multidisciplinariedade, Interdisciplinariedade e Transdisciplinariedade desde sua relação com a formação. Posteriormente, se identificam percepções de estudantes em processos de formação em Investigação mediante uma análise exploratório dos resultados obtidos de uma enquete de avaliação de uma atividade transdisciplinar denominada "foro de lições aprendidas", a análise se realizou mediante uma prova chi-quadrado com uma confiabilidade de 95% para uma amostra de 200 estudantes da faculdade de engenharia para o ano 2016. Resultados. Tendo em conta a rejeição de independência para 24 2 36,4, onde 24, representam os graus de liberdade com um erro de 5 % , se conserva a hipótese inicial. Conclusão. Existe independência entre semestre e o conceito dos estudantes sobre da sua formação por competências em relação ao foro, o qual implica à necessidade de propor estratégias de intervenção para a transgressão do disciplinar e potenciar o pensamento complexo nos estudantes.

Comparación clínica entre el tratamiento ortodóncico facilitado por corticotomía y ortodoncia convencional (estudio piloto) / Clinical comparison of orthodontic treatment facilitated by corticotomy and conventional orthodontics (pilot study)

El objetivo de este estudio fue comparar clínicamente la velocidad del movimiento ortodóncico y cambios en los parámetros periodontales en pacientes con ortodoncia convencional y ortodoncia facilitada con corticotomía para el tratamiento de apiñamiento dental anterior. Un total de 10 pacientes participaron en el estudio de la siguiente forma: 5 pacientes grupo ortodoncia y corticotomía (EXP) y 5 pacientes grupo ortodoncia (CONT). Se realizó examen clínico completo que incluyó análisis radiográfico, periodontal y modelos de estudio. Las mediciones sobre los cambios periodontal, tabla ósea bucal, movimiento lineal y angular fueron medidos al inicio, 30, 90 y 210 días posterior a los procedimientos. Las diferencias entre los grupos fueron establecidas con la prueba t-Student para muestras pareadas y no pareadas dependiendo del caso. Los dientes sometidos a ortodoncia y corticotomía mostraron una mayor velocidad del movimiento en comparación con el grupo control durante los primero 30 días de examinación (P<0,05). No hubo diferencias estadísticamente significativas en los parámetros clínicos periodontales y grosor de la tabla ósea bucal entre los grupos. En conclusión, la corticotomía acelera el movimiento ortodóncico durante los primero 30 días postquirúrgicos con lo cual se puede reducir el tiempo de tratamiento en pacientes con apiñamiento severo. Adicionalmente, los parámetros clínicos periodontales y el volumen de la tabla ósea bucal de los dientes sometidos a corticotomía se mantienen estables después del procedimiento.

The aim of this study was to compare clinically speed orthodontic movement and changes in periodontal parameters in patients with conventional orthodontics and orthodontics with corticotomy for the treatment of anterior dental crowding. A total of 10 patients participated in the study: 5 patients in the orthodontics and corticotomy group (EXP) and 5 in the orthodontics patient group (CONT). Complete clinical examination included radiographic, periodontal and study models analysis was carried out. Measurements on periodontal changes, buccal bone plate, linear and angular movement were measured at baseline at 30, 90 and 210 days after the procedures. Differences between groups were established with the t-Student for paired and unpaired samples depending on the case. Teeth undergoing orthodontic corticotomy showed greater speed of movement compared to the control group during the first 30 days examination (P <0.05). There were no statistically significant differences in periodontal clinical parameters and thickness of the buccal bone plate between the groups. In conclusion, accelerated corticotomy orthodontic movement during the first 30 days postoperative, can reduce the treatment period in patients with severe crowding. Additionally, clinical periodontal parameters and volume of the buccal bone plate teeth undergoing corticotomy remained stable after the procedure.

Vitamin D deficiency at pediatric intensive care admission / Deficiência de vitamina D em internações na unidade de terapia intensiva pediátrica

OBJECTIVE: to assess whether 25hydroxivitaminD or 25(OH)vitD deficiency has a high prevalence at pediatric intensive care unit (PICU) admission, and whether it is associated with increased prediction of mortality risk scores. METHOD: prospective observational study comparing 25(OH)vitD levels measured in 156 patients during the 12 hours after critical care admission with the 25(OH)vitD levels of 289 healthy children. 25(OH)vitD levels were also compared between PICU patients with pediatric risk of mortality III (PRISM III) or pediatric index of mortality 2 (PIM 2) > p75 [(group A; n = 33) vs. the others (group B; n = 123)]. Vitamin D deficiency was defined as < 20 ng/mL levels. RESULTS: median (p25-p75) 25(OH)vitD level was 26.0 ng/mL (19.2-35.8) in PICU patients vs. 30.5 ng/mL (23.2-38.6) in healthy children (p = 0.007). The prevalence of 25(OH)vitD < 20 ng/mL was 29.5% (95% CI: 22.0-37.0) vs. 15.6% (95% CI: 12.2-20.0) (p = 0.01). Pediatric intensive care patients presented an odds ratio (OR) for hypovitaminosis D of 2.26 (CI 95%: 1.41-3.61). 25(OH)vitD levels were 25.4 ng/mL (CI 95%: 15.5-36.0) in group A vs. 26.6 ng/mL (CI 95%: 19.3-35.5) in group B (p = 0.800). CONCLUSIONS: hypovitaminosis D incidence was high in PICU patients. Hypovitaminosis D was not associated with higher prediction of risk mortality scores. .

OBJETIVO: avaliar se a deficiência da 25-hidroxivitamina D, ou 25 (OH) vitD, tem prevalência elevada em internações na unidade de terapia intensiva pediátrica, e se estaria relacionada à previsão de escores de risco de mortalidade. MÉTODO: estudo observacional prospectivo comparando níveis de 25 (OH) vitD de 156 pacientes, mensurados nas primeiras 12 horas da internação em terapia intensiva, com níveis de 25 (OH) vitD de 289 crianças saudáveis. Os níveis de 25 (OH) vitD também foram comparados entre pacientes na UTIP com escore PRISM III ou PIM 2 > p75 (Grupo A; n = 33), e o restante, (Grupo B; n = 123). A deficiência de vitamina D foi definida como níveis < 20 ng/mL. RESULTADOS: o nível médio (p25-p75) de 25 (OH) vitD foi 26,0 ng/mL (19,2-35,8) em pacientes internados na UTIP, em comparação a 30,5 ng/mL (23,2-38,6) em crianças saudáveis (p = 0,007). A prevalência de 25 (OH) vitD < 20 ng/mL foi de 29,5% (IC 95%, 22,0-37,0), em comparação a 15,6% (IC 95%,12,2-20,0) (p = 0,01). Os pacientes em terapia intensiva pediátrica apresentaram uma razão de chance (RC) para hipovitaminose D de 2,26 (IC 95%, 1,41-3,61). Os níveis de 25 (OH) vitD foram 25,4 ng/mL (IC 95%, 15,5-36,0) no grupo A, em comparação a 26,6 ng/mL (IC 95%, 19,3-35,5) no grupo B (p = 0,800). CONCLUSÕES: a incidência de hipovitaminose D foi elevada em pacientes em terapia intensiva pediátrica, mas não foi associada à maior previsão de escores de risco de mortalidade. .

Vitamin D deficiency at pediatric intensive care admission.

OBJECTIVE: to assess whether 25hydroxivitaminD or 25(OH)vitD deficiency has a high prevalence at pediatric intensive care unit (PICU) admission, and whether it is associated with increased prediction of mortality risk scores. METHOD: prospective observational study comparing 25(OH)vitD levels measured in 156 patients during the 12 hours after critical care admission with the 25(OH)vitD levels of 289 healthy children. 25(OH)vitD levels were also compared between PICU patients with pediatric risk of mortality III (PRISM III) or pediatric index of mortality 2 (PIM 2) > p75 [(group A; n = 33) vs. the others (group B; n = 123)]. Vitamin D deficiency was defined as < 20 ng/mL levels. RESULTS: median (p25-p75) 25(OH)vitD level was 26.0 ng/mL (19.2-35.8) in PICU patients vs. 30.5 ng/mL (23.2-38.6) in healthy children (p = 0.007). The prevalence of 25(OH)vitD < 20 ng/mL was 29.5% (95% CI: 22.0-37.0) vs. 15.6% (95% CI: 12.2-20.0) (p = 0.01). Pediatric intensive care patients presented an odds ratio (OR) for hypovitaminosis D of 2.26 (CI 95%: 1.41-3.61). 25(OH)vitD levels were 25.4 ng/mL (CI 95%: 15.5-36.0) in group A vs. 26.6 ng/mL (CI 95%: 19.3-35.5) in group B (p = 0.800). CONCLUSIONS: hypovitaminosis D incidence was high in PICU patients. Hypovitaminosis D was not associated with higher prediction of risk mortality scores.

Estrés laboral en auxiliares de enfermería, Medellín-Colombia, 2011: Estrés laboral en auxiliares de enfermería / Job stress in nursing assistants, Medellín-Colombia, 2011

Objetivo: Identificar la presencia de estrés laboral en auxiliares de enfermería que trabajan en salas de hospitalización de adultos y las condiciones institucionales que lo configuran. Metodología: Estudio cualitativo de caso colectivo que parte de la determinación de características comunes o no entre los datos obtenidos de los casos individuales para su posterior análisis. Participaron seis casos: cinco mujeres y un hombre, auxiliares de enfermería con menos de 10 años de experiencia laboral en salas de hospitalización de adultos. Se utilizó la entrevista semiestructurada y se aplicó el consentimiento informado. Resultados: Las condiciones que provocaron el estrés fueron la Jornada laboral con tiempos reales de 13 ó 14 horas seguidas, sin espacios para la alimentación y el descanso, y actividades adicionales que no se encuentran dentro de las funciones normatizadas pero que son delegadas por otros profesionales de la salud y que se vuelven una carga más para ellas, tales como el transporte de pacientes, apoyo durante transfusiones sanguíneas, limpieza del carro de medicamentos y el estar ahí dando cuidado permanente a los pacientes. Como aminorante del estrés aparecen las relaciones entre el equipo de trabajo. Conclusión: Se identifica la presencia del estrés laboral en las auxiliares de enfermería que trabajan en salas de hospitalización de adultos. Las condiciones descritas limitan el tiempo para el cuidado de los pacientes y el cuidado de sí mismas y alteran su condición física, psicológica y emocional, factores que se relacionan con el estrés laboral. Salud UIS 2013; 45 (1): 23-31.

Objective: To identify the presence of stress in nursing assistants working in adult hospital rooms and institutional conditions that shape it. Methodology: Qualitative study of collective case, it parts on the determination of common or not characteristics between obtained dates of individual cases for later analysis. Six participant cases composed by five female and one male nursing assistants with less than 10 years of experience in adults hospital rooms. A semi-structured interview and a informed consent was applied. Results: The conditions that mediate stress were the workday with times of 13 or 14 hours continues without spaces for eating and resting and additional activities which aren’t in theirs functions but are delegated by other health professionals that increase their workload, like a moving patients, supporting blood’s transfusions, medication cart cleansing, and giving permanently care to patients. The relationships among the team appear as buffering of stress. Conclusion: The presence of occupational stress in nursing assistants that work in adults hospital rooms was identified. The conditions described limits the time for patient care and self care and affects their physical, psychological and emotional conditions, factors related to job stress. Salud UIS 2013; 45 (1): 23-31.