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Fetal hemoglobin (HbF) reactivation expression through CRISPR-Cas9 is a promising strategy for the treatment of sickle cell disease (SCD). Here, we describe a genome editing strategy leading to reactivation of HbF expression by targeting the binding sites (BSs) for the lymphoma-related factor (LRF) repressor in the γ-globin promoters. CRISPR-Cas9 treatment in healthy donor (HD) and patient-derived HSPCs resulted in a high frequency of LRF BS disruption and potent HbF synthesis in their erythroid progeny. LRF BS disruption did not impair HSPC engraftment and differentiation but was more efficient in SCD than in HD cells. However, SCD HSPCs showed a reduced engraftment and a myeloid bias compared with HD cells. We detected off-target activity and chromosomal rearrangements, particularly in SCD samples (likely because of the higher overall editing efficiency) but did not impact the target gene expression and HSPC engraftment and differentiation. Transcriptomic analyses showed that the editing procedure results in the up-regulation of genes involved in DNA damage and inflammatory responses, which was more evident in SCD HSPCs. This study provides evidence of efficacy and safety for an editing strategy based on HbF reactivation and highlights the need of performing safety studies in clinically relevant conditions, i.e., in patient-derived HSPCs.
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Physical activity and sports can be considered important sources of motivation for people with visual impairments; however limited attention has been given to blind peoples' involvement in sports and to the intra-personal variables such as self-esteem and motivation that affect their sport participation. The purpose of the present study was a) to explore the relationship between self-esteem and intrinsic motivation in blind athletes, and b) to assess the levels of self-esteem and intrinsic motivation and identify differences in intrinsic motivation and self-esteem as a function of gender and sport type (individual-team sports). Participants were 78 (27 females) Greek blind athletes (30.29 ± 1.18 years of age) competing in team (N = 44) and individual (N = 34) sports. Participants completed the Intrinsic Motivation Inventory questionnaire (IMI) and Rosenberg Self-Esteem Scale (RSES) questionnaire. The findings revealed that high values on self-esteem could predict blind athletes' perceived competence (p = .001) and strengthen their intrinsic motivation while enabling them to regulate pressure and tension (p < .05). However, female athletes appeared to be more pressured than males (p < .05), and interestingly, team sport athletes scored higher in intrinsic motivation than individual sports athletes (p < .05). The findings provide useful insights regarding the role of self-esteem in blind athletes' motivational profiles and draw attention to the role of perceived competence in blind athletes' sporting experiences, emphasizing that team sport context may serve as a fertile ground for enhancing intrinsic motivation.
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Introduction: The expansive curricular volume of healthcare education makes a necessity the incorporation of innovative methods and immersive media in it. The core challenge in such approaches is the timely development of relevant immersive content such as Virtual, Augmented or Mixed Reality (VR/AR/MR) resources for healthcare topics. There is currently significant interest in the use of co-creative methods for streamlining immersive content development. Methods: A core research pursuit in this translational research field is the formulation of evidence-based, optimized workflows that streamline immersive content creation allowing for rapid expansion of innovative educational approaches in healthcare curricula. The purpose of this paper is to aggregate the perceptions of healthcare technologists and educators who participated in a series of co-creation sessions in order to elicit their best practice insights for design and development of XR educational resources using co-creative methods. Results: According to our thematic analysis, findings of the qualitative study demonstrated that a rigorous organizational approach is required to maintain a constructive exchange of information and to keep the design process alive for both content and technical experts. In addition, rapid prototype and display of co-created features can empower their contributions and help them design more efficiently. Discussion: Co-creative content production can benefit from adaption of existing frameworks and lightweight authoring environments that can facilitate generalized XR content development use cases.
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In this guide we provide instructions and recommendations about creating and running escape rooms for healthcare education. In recent years there has been a growing interest in adopting escape rooms as an educational tool to be included in healthcare curricula, and we attempt to explain why and how these tools are fit for the particularities of this type of education. We first describe the steps that a design team will have to follow to create an educational escape room from scratch, from core characteristics like target audience and learning goals to actual puzzle design and testing. We then continue by providing recommendations to operators and lecturers about how to run such escape room as part of an overall teaching session that also includes a lecture, briefing, debriefing and evaluation. We finalise this guide by listing a set of tools for validating and evaluating these types of escape rooms.
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BACKGROUND: Atrial fibrillation (AF) is the most common sustained cardiac arrhythmia and is a leading cause of mortality and morbidity. Patient knowledge about AF and its management is paramount but often limited. Patients need to be appropriately informed about treatment options, medicinal adherence, and potential consequences of nonadherence, while also understanding treatment goals and expectations from it. Mobile health apps have experienced an explosion both in their availability and acceptance as "soft interventions" for patient engagement and education; however, the prolific nature of such solutions revealed a gap in the evidence base regarding their efficacy and impact. Virtual patients (VPs), interactive computer simulations, have been used as learning activities in modern health care education. VPs demonstrably improved cognitive and behavioral skills; hence, they have been effectively implemented across undergraduate and postgraduate curricula. However, their application in patient education has been rather limited so far. OBJECTIVE: This work aims to implement and evaluate the efficacy of a mobile-deployed VP regimen for the education and engagement of patients with AF on crucial topics regarding their condition. A mobile VP app is being developed with the goal of each VP being a simple scenario with a set goal and very specific messages and will be subsequently attempted and evaluated. METHODS: A mobile VP player app is being developed so as to be used for the design of 3 educational scenarios for AF management. A pseudorandomized controlled trial for the efficacy of VPs is planned to be executed at 3 sites in Greece, Ukraine, and Kazakhstan for patients with AF. The Welch t test will be used to demonstrate the performance of patients' evaluation of the VP experience. RESULTS: Our study is at the development stage. A preliminary study regarding the system's development and feasibility was initiated in December 2022. The results of our study are expected to be available in 2024 or when the needed sample size is achieved. CONCLUSIONS: This study aims to evaluate and demonstrate the first significant evidence for the value of VP resources in outreach and training endeavors for empowering and patients with AF and fostering healthy habits among them. INTERNATIONAL REGISTERED REPORT IDENTIFIER (IRRID): PRR1-10.2196/45946.
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ENTICE aimed to use co-creative methodologies in order to build a solid creation pipeline for medical experiential content. The project has developed and evaluated immersive learning resources and tools aiming to support well-defined learning objectives using tangible and intangible resources (AR/VR/MR, 3D printing) that are highly sought in the fields of anatomy and surgery. In this paper the preliminary results from the evaluation of the learning resources and tools in 3 countries as well as the lessons learnt are presented towards to the improvement of the medical education process.
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Educación Médica , Realidad Virtual , Aprendizaje , Terapia Conductista , Impresión TridimensionalRESUMEN
Virtual reality (VR) technologies have rapidly developed in the past few years. The most common application of the technology, apart from gaming, is for educational purposes. In the field of healthcare, VR technologies have been applied in several areas. Among them is surgical education. With the use of VR, surgical pathways along with the training of surgical skills can be explored safely, in a cost-effective manner. The aim of this mini-review was to explore the use of VR in surgical education and in the 3D reconstruction of internal organs and viable surgical pathways. Finally, based on the outcomes of the included studies, an ecosystem for the implementation of surgical training was proposed.
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BACKGROUND: This article presents the system architecture and validation of the NeuroSuitUp body-machine interface (BMI). The platform consists of wearable robotics jacket and gloves in combination with a serious game application for self-paced neurorehabilitation in spinal cord injury and chronic stroke. METHODS: The wearable robotics implement a sensor layer, to approximate kinematic chain segment orientation, and an actuation layer. Sensors consist of commercial magnetic, angular rate and gravity (MARG), surface electromyography (sEMG), and flex sensors, while actuation is achieved through electrical muscle stimulation (EMS) and pneumatic actuators. On-board electronics connect to a Robot Operating System environment-based parser/controller and to a Unity-based live avatar representation game. BMI subsystems validation was performed using exercises through a Stereoscopic camera Computer Vision approach for the jacket and through multiple grip activities for the glove. Ten healthy subjects participated in system validation trials, performing three arm and three hand exercises (each 10 motor task trials) and completing user experience questionnaires. RESULTS: Acceptable correlation was observed in 23/30 arm exercises performed with the jacket. No significant differences in glove sensor data during actuation state were observed. No difficulty to use, discomfort, or negative robotics perception were reported. CONCLUSIONS: Subsequent design improvements will implement additional absolute orientation sensors, MARG/EMG based biofeedback to the game, improved immersion through Augmented Reality and improvements towards system robustness.
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Dispositivo Exoesqueleto , Rehabilitación Neurológica , Robótica , Rehabilitación de Accidente Cerebrovascular , Accidente Cerebrovascular , Dispositivos Electrónicos Vestibles , HumanosRESUMEN
Beta-like globin gene expression is developmentally regulated during life by transcription factors, chromatin looping and epigenome modifications of the ß-globin locus. Epigenome modifications, such as histone methylation/demethylation and acetylation/deacetylation and DNA methylation, are associated with up- or down-regulation of gene expression. The understanding of these mechanisms and their outcome in gene expression has paved the way to the development of new therapeutic strategies for treating various diseases, such as ß-hemoglobinopathies. Histone deacetylase and DNA methyl-transferase inhibitors are currently being tested in clinical trials for hemoglobinopathies patients. However, these approaches are often uncertain, non-specific and their global effect poses serious safety concerns. Epigenome editing is a recently developed and promising tool that consists of a DNA recognition domain (zinc finger, transcription activator-like effector or dead clustered regularly interspaced short palindromic repeats Cas9) fused to the catalytic domain of a chromatin-modifying enzyme. It offers a more specific targeting of disease-related genes (e.g., the ability to reactivate the fetal γ-globin genes and improve the hemoglobinopathy phenotype) and it facilitates the development of scarless gene therapy approaches. Here, we summarize the mechanisms of epigenome regulation of the ß-globin locus, and we discuss the application of epigenome editing for the treatment of hemoglobinopathies.
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Epigénesis Genética , Hemoglobinopatías , Humanos , Epigénesis Genética/genética , Epigenoma , Hemoglobinopatías/genética , Hemoglobinopatías/terapia , Globinas beta/genética , Globinas beta/metabolismo , Cromatina , ADN/metabolismoRESUMEN
ß-Thalassemia (BT) is one of the most common genetic diseases worldwide and is caused by mutations affecting ß-globin production. The only curative treatment is allogenic hematopoietic stem/progenitor cells (HSPCs) transplantation, an approach limited by compatible donor availability and immunological complications. Therefore, transplantation of autologous, genetically-modified HSPCs is an attractive therapeutic option. However, current gene therapy strategies based on the use of lentiviral vectors are not equally effective in all patients and CRISPR/Cas9 nuclease-based strategies raise safety concerns. Thus, base editing strategies aiming to correct the genetic defect in patients' HSPCs could provide safe and effective treatment. Here, we developed a strategy to correct one of the most prevalent BT mutations (IVS1-110 [G>A]) using the SpRY-ABE8e base editor. RNA delivery of the base editing system was safe and led to â¼80% of gene correction in the HSPCs of patients with BT without causing dangerous double-strand DNA breaks. In HSPC-derived erythroid populations, this strategy was able to restore ß-globin production and correct inefficient erythropoiesis typically observed in BT both in vitro and in vivo. In conclusion, this proof-of-concept study paves the way for the development of a safe and effective autologous gene therapy approach for BT.
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Talasemia beta , Humanos , Talasemia beta/genética , Talasemia beta/terapia , Edición Génica , Sistemas CRISPR-Cas , Mutación , Globinas beta/genéticaRESUMEN
The purpose was to assess the effect of an 11-week resisted swim training program using a water parachute on the inter-arm coordination in front crawl swimming. Fourteen female swimmers were divided into a control and an experimental group, and followed the same training program, except for the use of a water parachute by the experimental group in specific sprint sets. Before and after the intervention, the underwater motion of the swimmers was recorded using four camcorders and the digitisation process was undertaken manually. The two-way repeated measures analyses of variance revealed a significant increase of the swimming velocity, due to a significant increased stroke rate, because of the significant reduction of the total stroke duration, only in the experimental group. Moreover, their absolute and relative duration of the propulsive pull and push phases increased significantly, while the absolute and relative duration of their non-propulsive entry & catch and recovery phases decreased significantly. These modifications caused a significant increase in the index of coordination. Thus, this 11-week sprint resisted intervention program seems to improve the propulsion continuity and could be considered as an effective form of training, increasing the stroke rate and consequently the swimming velocity.
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Natación , Agua , Humanos , Femenino , Fenómenos Biomecánicos , Movimiento (Física)RESUMEN
Sickle cell disease and ß-thalassemia affect the production of the adult ß-hemoglobin chain. The clinical severity is lessened by mutations that cause fetal γ-globin expression in adult life (i.e., the hereditary persistence of fetal hemoglobin). Mutations clustering ~200 nucleotides upstream of the HBG transcriptional start sites either reduce binding of the LRF repressor or recruit the KLF1 activator. Here, we use base editing to generate a variety of mutations in the -200 region of the HBG promoters, including potent combinations of four to eight γ-globin-inducing mutations. Editing of patient hematopoietic stem/progenitor cells is safe, leads to fetal hemoglobin reactivation and rescues the pathological phenotype. Creation of a KLF1 activator binding site is the most potent strategy - even in long-term repopulating hematopoietic stem/progenitor cells. Compared with a Cas9-nuclease approach, base editing avoids the generation of insertions, deletions and large genomic rearrangements and results in higher γ-globin levels. Our results demonstrate that base editing of HBG promoters is a safe, universal strategy for treating ß-hemoglobinopathies.
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Anemia de Células Falciformes , Talasemia beta , Humanos , Hemoglobina Fetal/genética , Hemoglobina Fetal/metabolismo , gamma-Globinas/genética , Talasemia beta/genética , Talasemia beta/terapia , Anemia de Células Falciformes/genética , Células Madre Hematopoyéticas/metabolismoRESUMEN
BACKGROUND: Spinal cord injury (SCI) constitutes a major sociomedical problem, impacting approximately 0.32-0.64 million people each year worldwide; particularly, it impacts young individuals, causing long-term, often irreversible disability. While effective rehabilitation of patients with SCI remains a significant challenge, novel neural engineering technologies have emerged to target and promote dormant neuroplasticity in the central nervous system. OBJECTIVE: This study aims to develop, pilot test, and optimize a platform based on multiple immersive man-machine interfaces offering rich feedback, including (1) visual motor imagery training under high-density electroencephalographic recording, (2) mountable robotic arms controlled with a wireless brain-computer interface (BCI), (3) a body-machine interface (BMI) consisting of wearable robotics jacket and gloves in combination with a serious game (SG) application, and (4) an augmented reality module. The platform will be used to validate a self-paced neurorehabilitation intervention and to study cortical activity in chronic complete and incomplete SCI at the cervical spine. METHODS: A 3-phase pilot study (clinical trial) was designed to evaluate the NeuroSuitUp platform, including patients with chronic cervical SCI with complete and incomplete injury aged over 14 years and age-/sex-matched healthy participants. Outcome measures include BCI control and performance in the BMI-SG module, as well as improvement of functional independence, while also monitoring neuropsychological parameters such as kinesthetic imagery, motivation, self-esteem, depression and anxiety, mental effort, discomfort, and perception of robotics. Participant enrollment into the main clinical trial is estimated to begin in January 2023 and end by December 2023. RESULTS: A preliminary analysis of collected data during pilot testing of BMI-SG by healthy participants showed that the platform was easy to use, caused no discomfort, and the robotics were perceived positively by the participants. Analysis of results from the main clinical trial will begin as recruitment progresses and findings from the complete analysis of results are expected in early 2024. CONCLUSIONS: Chronic SCI is characterized by irreversible disability impacting functional independence. NeuroSuitUp could provide a valuable complementary platform for training in immersive rehabilitation methods to promote dormant neural plasticity. TRIAL REGISTRATION: ClinicalTrials.gov NCT05465486; https://clinicaltrials.gov/ct2/show/NCT05465486. INTERNATIONAL REGISTERED REPORT IDENTIFIER (IRRID): PRR1-10.2196/41152.
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Considerable attention nowadays has been given to collaborative learning as a powerful way of learning, however, the ability to provide student-centered learning, meet student needs and provide them with the opportunity to learn, is a challenging task and has received significant research attention. CLUEDUP proposes an innovative educational protocol that integrates game-based learning with pedagogical approaches that matches students' needs and skills and promotes collaborative interactive and decision-based learning.
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Prácticas Interdisciplinarias , Humanos , Aprendizaje , EstudiantesRESUMEN
Motor neuron diseases are untreatable with common pharmacological approaches. Spinal muscular atrophy (SMA) is caused by SMN1 gene mutations leading to lowered SMN expression. Symptoms are alleviated in infants with a higher copy number of the SMN2 gene, which, however, displays a splicing defect resulting in low SMN levels. Amyotrophic lateral sclerosis (ALS) is caused by a number of mutations, with C9orf72 repeat expansions the most common genetic cause and SOD1 gain-of-function mutations the first genetic cause identified for this disease. Genetic therapies based on oligonucleotides that enhance SMN2 splicing and SMN production or lower SOD1 expression have shown promise in initial clinical trials for individuals with SMA and ALS harboring SOD1 mutations, respectively. Gene addition/silencing approaches using adeno-associated viruses (AAVs) are also currently under clinical investigation in trials for SMA and ALS. Here we provide a brief overview of these efforts and their advantages and challenges. We also review genome editing approaches aimed at correcting the disease-causing mutations or modulating the expression of genetic modifiers, e.g., by repairing SOD1 mutations or the SMN2 splicing defect or deleting C9orf72 expanded repeats. These studies have shown promising results to approach therapeutic trials that should significantly lower the progression of these deadly disorders.
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Esclerosis Amiotrófica Lateral , Atrofia Muscular Espinal , Esclerosis Amiotrófica Lateral/genética , Esclerosis Amiotrófica Lateral/metabolismo , Esclerosis Amiotrófica Lateral/terapia , Edición Génica , Humanos , Lactante , Atrofia Muscular Espinal/genética , Atrofia Muscular Espinal/metabolismo , Atrofia Muscular Espinal/terapia , Oligonucleótidos/metabolismo , Empalme del ARN , Proteína 1 para la Supervivencia de la Neurona Motora/genética , Proteína 1 para la Supervivencia de la Neurona Motora/metabolismoRESUMEN
Nuclease-based genome editing strategies hold great promise for the treatment of blood disorders. However, a major drawback of these approaches is the generation of potentially harmful double strand breaks (DSBs). Base editing is a CRISPR-Cas9-based genome editing technology that allows the introduction of point mutations in the DNA without generating DSBs. Two major classes of base editors have been developed: cytidine base editors or CBEs allowing C>T conversions and adenine base editors or ABEs allowing A>G conversions. The scope of base editing tools has been extensively broadened, allowing higher efficiency, specificity, accessibility to previously inaccessible genetic loci and multiplexing, while maintaining a low rate of Insertions and Deletions (InDels). Base editing is a promising therapeutic strategy for genetic diseases caused by point mutations, such as many blood disorders and might be more effective than approaches based on homology-directed repair, which is moderately efficient in hematopoietic stem cells, the target cell population of many gene therapy approaches. In this review, we describe the development and evolution of the base editing system and its potential to correct blood disorders. We also discuss challenges of base editing approaches-including the delivery of base editors and the off-target events-and the advantages and disadvantages of base editing compared to classical genome editing strategies. Finally, we summarize the recent technologies that have further expanded the potential to correct genetic mutations, such as the novel base editing system allowing base transversions and the more versatile prime editing strategy.
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In the present study, a buckling analysis of laminated composite rectangular plates reinforced with multiwalled carbon nanotube (MWCNT) inclusions is carried out using the finite element method (FEM). The rule of mixtures and the Halpin-Tsai model are employed to calculate the elastic modulus of the nanocomposite matrix. The effects of three critical factors, including random dispersion, waviness, and agglomeration of MWCNTs in the polymer matrix, on the material properties of the nanocomposite are analyzed. Then, the critical buckling loads of the composite plates are numerically determined for different design parameters, such as plate side-to-thickness ratio, elastic modulus ratio, boundary conditions, layup schemes, and fiber orientation angles. The influence of carbon nanotube fillers on the critical buckling load of a nanocomposite rectangular plate, considering the modified Halpin-Tsai micromechanical model, is demonstrated. The results are in good agreement with experimental and other theoretical data available in the open literature.
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BACKGROUND: The role of emotion is crucial to the learning process, as it is linked to motivation, interest, and attention. Affective states are expressed in the brain and in overall biological activity. Biosignals, like heart rate (HR), electrodermal activity (EDA), and electroencephalography (EEG) are physiological expressions affected by emotional state. Analyzing these biosignal recordings can point to a person's emotional state. Contemporary medical education has progressed extensively towards diverse learning resources using virtual reality (VR) and mixed reality (MR) applications. OBJECTIVE: This paper aims to study the efficacy of wearable biosensors for affect detection in a learning process involving a serious game in the Microsoft HoloLens VR/MR platform. METHODS: A wearable array of sensors recording HR, EDA, and EEG signals was deployed during 2 educational activities conducted by 11 participants of diverse educational level (undergraduate, postgraduate, and specialist neurosurgeon doctors). The first scenario was a conventional virtual patient case used for establishing the personal biosignal baselines for the participant. The second was a case in a VR/MR environment regarding neuroanatomy. The affective measures that we recorded were EEG (theta/beta ratio and alpha rhythm), HR, and EDA. RESULTS: Results were recorded and aggregated across all 3 groups. Average EEG ratios of the virtual patient (VP) versus the MR serious game cases were recorded at 3.49 (SD 0.82) versus 3.23 (SD 0.94) for students, 2.59 (SD 0.96) versus 2.90 (SD 1.78) for neurosurgeons, and 2.33 (SD 0.26) versus 2.56 (SD 0.62) for postgraduate medical students. Average alpha rhythm of the VP versus the MR serious game cases were recorded at 7.77 (SD 1.62) µV versus 8.42 (SD 2.56) µV for students, 7.03 (SD 2.19) µV versus 7.15 (SD 1.86) µV for neurosurgeons, and 11.84 (SD 6.15) µV versus 9.55 (SD 3.12) µV for postgraduate medical students. Average HR of the VP versus the MR serious game cases were recorded at 87 (SD 13) versus 86 (SD 12) bpm for students, 81 (SD 7) versus 83 (SD 7) bpm for neurosurgeons, and 81 (SD 7) versus 77 (SD 6) bpm for postgraduate medical students. Average EDA of the VP versus the MR serious game cases were recorded at 1.198 (SD 1.467) µS versus 4.097 (SD 2.79) µS for students, 1.890 (SD 2.269) µS versus 5.407 (SD 5.391) µS for neurosurgeons, and 0.739 (SD 0.509) µS versus 2.498 (SD 1.72) µS for postgraduate medical students. The variations of these metrics have been correlated with existing theoretical interpretations regarding educationally relevant affective analytics, such as engagement and educational focus. CONCLUSIONS: These results demonstrate that this novel sensor configuration can lead to credible affective state detection and can be used in platforms like intelligent tutoring systems for providing real-time, evidence-based, affective learning analytics using VR/MR-deployed medical education resources.