They were then subject to a further heat/cool cycle, which was monitored by simultaneous differential scanning calorimetry – X-ray diffraction to observe the phase changes occurring. These varied with the composition of the blend. Upon short-term storage, formulations with low trehalose contents  recrystallised into form I FFA, while higher trehalose contents crystallised to FFA form IV. When heated, all FFA trehalose combinations ultimately recrystallised into form I before melting. Upon a second cooling cycle, systems with low trehalose content  recrystallised into form IV, while higher trehalose contents led to FFA form I. It is thus clear that even with a single excipient it is possible to control the crystallisation pathway through judicious choice of the formulation parameters.The combination of poorly-soluble drugs with small molecule co-formers to generate amorphous solid dispersions (ASDs) has great potential to improve dissolution rate and kinetic solubility, and thus increase the bioavailability of these active ingredients. However, such ASDs are known to be unstable and to crystallise upon storage or heating. In this work, we explore the crystallisation of flufenamic acid (FFA) from ASDs prepared with trehalose. FFA-trehalose mixtures were prepared at a range of   One option to overcome this issue is to use amorphous systems . HoweverPolymers have been frequently employed to inhibit crystallisation and increase the stability of amorphous materials . An alteIn this work, we explore crystallisation and interpolymorph conversion in composites of flufenamic acid amino]benzoic acid) and trehalose. FFA is a non-steroidal anti-inflammatory drug (NSAID) which falls in Class II of the BCS and thus suffers from low solubility. To overcome this limitation, amorphization has been explored . FFA is w/w FFA-polymer blend the drug remained mostly amorphous upon heating. Both the viscosity and substitution pattern of HPMC are found to influence the recrystallisation process. It is thus clear that there are myriad and subtle effects involved in the recrystallisation from polymer-drug ASDs. Here, we sought to expand on our previous study to explore the effects of using small-molecule excipients for ASD preparation.We have previously shown that heating a glass of pure FFA results in an initial recrystallisation into an unknown polymorph at ca. 54 °C, and that this then converts to form IV and then to form I . The behd-glucopyranosyl-α-d-glucopyranoside). It has a high glass transition temperature, and is widely utilised in the food and pharmaceutical industries  and trehalose  were purchased from Acros.2.22.2.1w/w; FFA:trehalose). The samples were then mixed for ca. 3 min using a vortex mixer. Ca. 5–10 mg of sample was heated from 0 °C to 150 °C at 10 °C/min and then cooled to 0 °C in a TA Instruments Q2000 DSC to produce amorphous solid dispersions. The obtained solids were stored in a fridge (6 °C) before characterisation by DSC-XRD. DSC measurements were performed with a modified Q20 DSC (TA Instruments), with the instrument modification, experiment set up and data analysis protocols being the same as described previously  were collected using the Diamond Generic Data Acquisition (GDA) software. The resultant two-dimensional Pixium images were masked, and processed using the DAWN Science Workbench software into one-dimensional diffraction patterns by azimuthal integration . TOPAS-A2.2.2−1 with 64 scans at a resolution of 4 cm−1. FFA, trehalose and the samples explored in DSC-XRD were all evaluated. The drug/trehalose mixtures were heated in the Q2000 DSC from 40 °C to 150 °C at 10 °C/min before equilibrating back to 0 °C and then reheated to the temperature of interest  at 10 °C/min. The samples were then removed from the DSC and characterised by FTIR.FTIR spectroscopy was conducted with a Perkin Elmer Spectrum 100 instrument. All spectra were recorded between 650 and 4000 cm3FFA/trehalose (FFA/T) blends were heated to melting and then cooled to 0 °C in a Q2000 DSC to produce amorphous materials. The DSC traces suggest that no recrystallisation took place during the cooling processes . The solids obtained were stored in the fridge before characterisation by DSC-XRD at the Diamond Light Source (< 5 days later). This was expected to minimise the extent of recrystallisation. However, from the XRD results it is clear that there are some reflections in the contour plots at the beginning of the DSC-XRD experiments, which suggests some degree of recrystallisation occurred during storage.3.1w/w) are presented in DSC-XRD data for a 1:2 FFA:trehalose weight ratio mixture  and 75 °C (during cooling)  using the Rietveld method. The pattern recorded at 78 °C comprises FFA form IV and trehalose β. This suggests FFA in the sample recrystallised into form IV during storage, and trehalose crystallised into its β form. Refinement against the pattern at 115 °C confirms that the solid is FFA form I and trehalose β. The pattern recorded at 121 °C during cooling is entirely trehalose β. Thus, during heating, FFA converts from form IV to form I and then form I melts at 135 °C. Trehalose remains in its β form throughout the experiment. During cooling, Rietveld refinement reveals that the pattern collected at 75 °C is FFA form I and trehalose β, which means in this formulation molten FFA recrystallises into form I upon cooling.In order to explore the presence of the different FFA polymorphs as a function of temperature, batch refinement was carried out and the results are plotted in −1 and 0.94 °C−1 respectively, with form I growing in more slowly then form IV disappears. During heating, some form IV converts to form I through a polymorphic transiton, while some becomes amorphous. The conversion of crystalline material to an amorphous phase clearly occurs more rapidly than the molecular rearrangements needed to form I. Thus, the decay of form IV happens faster than the growth of form I. The growth of form I peaks at 125 °C, after which the amount present decreases rapidly to a minimum at 145 °C. Both form I and IV stay at a minimum level before form I starts to grow back in during air cooling. The amount of form IV appears to increase at 140 °C, but this is an artefact arising because at this temperature FFA turns into a liquid. During batch refinement, the crystal structures of form I and IV are used as references to calculate the phase fractions and integral intensities. There is a fully amorphous diffraction pattern at temperatures above 140 °C, which means that it is not possible to effectively fit the structures to the datasets and thus the phase fractions calculated are not fully accurate.In 3.2w/w) are given in DSC-XRD data for the mixture with a FFA:trehalose ratio of 1:1 w/w  and 68 °C (cooling) were examined individually . The initial material (47 °C) comprises FFA form IV and trehalose β . As for the FFA/T 1: 2 w/w sample, FFA/T 1: 1 w/w recrystallised into FFA form IV and trehalose β during storage. The refinement against the pattern at 91 °C shows that the solid is a mixture of form I and form II FFA and trehalose β. The pattern recorded at 128 °C after the exothermic event comprises form I FFA and trehalose β. At 157 °C, the sample is identified as being entirely trehalose β. Thus, during heating FFA converts from form IV to form I/II. It appears that the form II generated subsequently converts to form I, which then melts at 135 °C while trehalose remains in its β form. Refinement at 68 °C, recorded on cooling, shows the sample to comprise FFA form I and trehalose β. Here again the FFA recrystallises into form I on cooling.Batch refinement was performed on the full set of XRD data, and −1 and 0.69 °C−1, respectively. Similarly to the 1:2 data in −1) and form II (0.32 °C−1) are almost the same, suggesting that form IV converts to form I and II at the same speed.In 3.3The mixture comprising FFA:trehalose 3:2 w/w shows the same phase transitions as the 1:2 w/w analogue . A smallPhase identifications were carried out with Rietveld refinement against patterns recorded at 71 °C, 120 °C, 160 °C (during heating) and 63 °C (during cooling). Analogous findings to those obtained with other FFA/T blends are observed. Selected XRD patterns and refinement parameters are shown in Fig. S4 and Table S3.−1 and 0.01 °C−1, respectively with form I growing more slowly. The phase fraction of form I at 100 °C (after the polymorphic transition) is somewhat lower than the maximum percentage of form IV, as previously suggesting that not all form IV converts to form I. During the cooling process, there are only form I appears in the sample.The relative proportions of FFA polymorphs were analysed by batch refinement . This re3.4w/w dispersion gives a different phase transition profile to the previous mixtures explored . This suggests that only a small amount of FFA crystallised at 87 °C, while most of remains in the amorphous state. Batch refinements were carried out to determine the amount of each FFA polymorph present in the sample as a function of temperature see . FFA in 3.5w/w), 100 and 120 °C (1:1 w/w). All the samples were heated in the DSC and then removed for FTIR characterisation.Drug-polymer interactions were studied by FTIR. Spectra were recorded for FFA and trehalose and compared with mixtures prepared as for DSC-XRD analysis and heated to 120 °C  scale" fill="currentColor" stroke="none"><path d="M0 440 l0 -40 480 0 480 0 0 40 0 40 -480 0 -480 0 0 -40z M0 280 l0 -40 480 0 480 0 0 40 0 40 -480 0 -480 0 0 -40z"/></g></svg>O and CC vibrations are at 1654 and 1578 cm−1, respectively , while the 1:1 w/w sample recrystallises into forms I and II at 100 °C and converts to form I at 120 °C. FTIR spectra are presented in ectively . For FFAectively b which s4w/w), FFA recrystallises into form I, while in the other formulations FFA form IV is generated upon storage. Before characterisation at Diamond, the samples were stored at 6 °C in a fridge, which suggests that a higher trehalose content can stabilise FFA form IV at low temperature.During heat-cool cycles in the DSC, FFA-trehalose mixtures give different phase transition behaviour, dependent on the relative amount of trehalose present. In the DSC-XRD study , all thew/w) recrystallised into form IV, while the FFA in the other formulations converted into form I. Since FTIR results suggest no H-bonds were formed between FFA and trehalose, this leads to the hypothesis that trehalose may not affect the relative phase stability of FFA, but it may alter the activation energy for the transformation of amorphous FFA to form I and form IV. Therefore, when the trehalose content is high enough, the activation energy for amorphous FFA to convert to form I will be low enough for this transition without passing through metastable form IV.During heating, phase transitions from form IV to other FFA forms occurred at around 80 °C for the FFA trehalose mixtures, as has also been reported for pure FFA . It thusThe literature shows polymers are able to stabilise FFA form IV or amorphous phases during heating ; however5w/w) recrystallised into form I FFA, while higher trehalose content led to FFA form IV being generated after storage. When heated in the DSC, all FFA trehalose mixtures ultimately led to form I FFA, though form II was also seen with a 1:1 w/w ratio. Upon a second cooling cycle, mixtures with low trehalose content  transformed into form IV, while FFA with higher trehalose content converted to form I. Different trehalose ratios to FFA ratios thus demonstrate the ability to influence the polymorphic transitions of FFA.Undertaking heat-cool cycles with FFA-trehalose mixtures prepared at different trehalose contents revealed that the phase transitions occurring varied with the composition of the blend. During storage, vitrified blends with low trehalose content (FFA: trehalose = 5:1 Conceptualization, Y·P. S.G., G.R.W.; methodology, Y.P., O.V.M.; formal analysis, Y.P.; investigation, Y.P., O.V.M.; resources, S.G., G.R.W.; data curation, Y.P, G.R.W.; writing—original draft preparation, Y.P.; writing—review and editing: Y.P., S.G., O.V.M., G.R.W.; visualization, Y.P.; supervision, S.G., G.R.W.; project administration, G.R.W.; funding acquisition, S.G., G.R.W. All authors have read and agreed to the published version of the manuscript.The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.


However, the abandonment rate of prosthetics is higher than 50% due to the high cost of rehabilitation. Virtual technology shows potential for improving the availability and cost-effectiveness of prosthetic rehabilitation. This article systematically reviews the application of virtual technology for the prosthetic rehabilitation of upper limb amputees. (2) Methods: We followed PRISMA review guidance, STROBE, and CASP to evaluate the included articles. Finally, 17 articles were screened from 22,609 articles. (3) Results: This study reviews the possible benefits of using virtual technology from four aspects: usability, flexibility, psychological affinity, and long-term affordability. Three significant challenges are also discussed: realism, closed-loop control, and multi-modality integration. (4) Conclusions: Virtual technology allows for flexible and configurable control rehabilitation, both during hospital admissions and after discharge, at a relatively low cost. The technology shows promise in addressing the critical barrier of current prosthetic training issues, potentially improving the practical availability of prosthesis techniques for upper limb amputees. Upper limb amputation poses many physical and psychological challenges that can significantly constrain one’s daily activities and occupational abilities ,2,3,4. FThe abandonment of upper limb prostheses can be attributed to several factors. First, despite intelligent prosthetic limbs providing a natural interaction between the user and machine, there were no significant improvements in self-reported functional recovery . ParticiIn contrast, computer-aided prosthetic training is a promising course of action for tackling the challenges associated with patient rehabilitation compliance, particularly those related to cost and motivation for training ,24,25. COverall, although the potential for virtual prosthetic rehabilitation exists as a viable, versatile, and cost-effective tool for amputation therapy, the extent of its effectiveness remains unclear. A systematic review is necessary to assess how to integrate this technology into traditional rehabilitation techniques to optimize its benefits. To obtain a more profound understanding of the therapeutic value of virtual environments for upper limb amputees, this study offers a comprehensive review of this topic and presents a broad discussion of the feasibility of virtual technology to enhance prosthetic rehabilitation. Firstly, we provide a detailed analysis of the theoretical applications of virtual technology to prosthetic rehabilitation. Secondly, we present a summary of current trends and future challenges using this technique. Our survey also seeks to identify gaps between the current research and clinical applications and propose potential avenues for future investigations in this field. The goal of this review is to contribute to the existing body of research by providing the benefits of virtual technology in upper limb rehabilitation and completing numerous investigations into its potential applications.The paper systematically evaluates a wide range of studies on prosthetic rehabilitation using computer-aided virtual technology. Given the plethora of available literature of many quantities and types, an approach with a more integrative and evidence-based perspective on upper limb amputation and associated prosthetic rehabilitation was used in this review. Accordingly, we adopted a five-stage framework for our review, as demonstrated by Whittemore and Knafl , consistThis review aims to critically evaluate the direct empirical evidence concerning the utilization of virtual technology in the prosthetic control rehabilitation of upper limb amputees. Furthermore, this review seeks to comprehensively analyze various approaches to upper limb amputation rehabilitation. The synthesis of the findings from the study offers valuable insights into the potential benefits of utilizing virtual technologies in overcoming the challenges associated with upper limb amputation. By exploring these approaches, all stakeholders could enhance their comprehension and refine upper limb rehabilitation strategies. This review facilitates the presentation and analysis of studies conducted within this particular scope. Ultimately, this research provides essential knowledge and guidance for practitioners and patients, enabling them to navigate the complexities of prosthetic rehabilitation and improve occupational and daily life outcomes for upper limb amputees.The study extensively searched electronic sources using various databases to identify relevant research studies , is examined by BN Perry et al.  as a traHowever, to realize an engaging experience for the control of virtual prostheses is still a formidable challenge faced by individuals with limb loss. The most important issue is how to establish a two-way interaction between the user and the virtual environment. To address this, Rahman Davoodi and Gerald E. Loeb  proposedEmpathy: The stakeholders engaged in the rehabilitation of upper limb amputees possess an intimate familiarity with the distinct requirements of individuals in varying circumstances based onthe virtual environment, thereby enabling the tailoring of appropriate training and rehabilitation regimens. Through direct observation of the rehabilitation needs of upper limb amputees, these stakeholders are able to conceive customized interventions tailored to address the specific challenges encountered by each individual. This personalized approach amplifies the efficacy of these training and rehabilitation activities. Consequently, the term “empathy” in this paper denotes the stakeholders’ capacity to comprehensively grasp the rehabilitative expectations stemming from the hardships endured by upper limb amputees, thereby fostering an empathetic connection with their rehabilitation needs.Hashet et al.  investigAdherence: Although various therapy techniques have been used, motivation remains a significant obstacle to the amount of training required for optimal recovery . VirtualAdaptability: Considering the potential adverse psychological effects of the virtual environment is also critical. Therefore, virtual approaches must be designed to enable adaptability depending on the mental states of users. Virtual technology can also modulate the difficulty, challenge, goal, feedback, interaction, and socialization of the training tasks to avoid possible frustration from amputees . Recent Budget constraints often lead to a lack of prioritization for new technologies, especially if current solutions are already beneficial. In a survey focused on physician attitudes toward new healthcare technology, it was found that 80% of respondents identified a lack of financial support as the main barrier . HoweverLow-cost: Budgetary constraints often lead to a decreased importance placed on innovative technologies, particularly when existing solutions are deemed sufficient in delivering benefits . The virVirtual technology offers numerous cost-saving benefits for upper limb amputees. First, unlike traditional prostheses, whose hardware items require frequent adjustment or replacement, the adaptability of virtual technology allows for software customization, eliminating the costs of physical components. Second, virtual technology can integrate realistic and haptic simulations to closely replicate the functionality of a natural limb. This immersive experience prompts more effective training with less rehabilitation time, potentially reducing the associated costs. Third, virtual training allows for remote monitoring and telerehabilitation, where healthcare professionals can track progress, offer guidance, and make necessary adjustments remotely, reducing the need for in-person visits and saving travel expenses. In conclusion, the adaptability of hardware, the ability to mimic natural limb functioning, and the potential for remote monitoring and telerehabilitation make virtual technology a more cost-effective solution for upper limb amputees compared to traditional prosthetic items, resulting in significant long-term cost savings.We conducted a cost-effectiveness analysis for the benefits of implementing virtual technology in motor rehabilitation compared with traditional methods. Specifically, we sought to determine if virtual approaches are more economically advantageous for all parties involved. We focused on stroke patients who also need motor function rehabilitation due to a lack of available data on prosthesis rehabilitation. From the patient’s perspective, traditional physical therapy services incur an annual cost of USD 11,689 per patient . In contFree software: Virtual technology with open-source software can be leveraged to offer a free solution for crafting virtual environments. Prominent examples of such software are Blender and Unity ,71, whicNE Bunderson  discusseBy tailoring the intensity of the virtual workouts to the patient’s specific capabilities and progress, therapists can potentially leverage these environments to contribute positively to the patient’s rehabilitation progress. Consequently, creating virtual environments using open-source software presents a promising avenue for advancing rehabilitative care for this population.In this study, we analyze factors that can contribute to increased patient engagement and improved adherence to prosthetic rehabilitation training through virtual technology. These factors form the basis of practical benefits for virtual therapy in amputee rehabilitation. Our process draws on the following four characteristics of the virtual environment: usability for prosthesis training, flexibility for the training platform, an affinity for user experience, and affordability for training costs.Notably, the factors extracted from our findings can potentially inform the future design and development of virtual systems for rehabilitation training. However, several challenges must be addressed to fully realize the potential of virtual technology in amputee rehabilitation . These iChallenges:Advances in virtual prosthetic training-based prosthesis rehabilitation are predominantly limited to academic research settings, with few practical applications in clinical environments.We identified a need for more research focused on incorporating closed-loop control of user feedback  into immersive virtual environments.One significant challenge to the widespread adoption of the technology discussed in this article lies in the limited number of amputee subjects involved in the experimental trials. Such trials pose challenges in obtaining generalizable results. As scientific knowledge advances, it becomes increasingly crucial to incorporate user-centered designs and larger sample sizes in studies related to clinical deployments. Subsequently, it becomes necessary to evaluate whether remote training options can support these studies.The elderly population may encounter challenges when using virtual rehabilitation technology. First, they may have deficiencies in the fundamental skills required for operating smart electronic devices. Second, the deterioration of neuromuscular function may result in the inability to operate the virtual human–machine system.Directions:Improving realism: This can be achieved by integrating novel technologies  and using realistic visual cues in simulated tasks. In addition, there is a need to enhance closed-loop control to improve the accuracy of motion tracking and achieve more natural movements in virtual settings. Improvement of virtual reality environments and virtual limbs in rehabilitation paradigms has been proposed to enhance their efficacy. The modeling of virtual limbs with a higher degree of realism and movement resolution has the potential to yield substantial benefits. In addition, future research should evaluate the impact of pre-prosthetic training methods on various outcomes in amputee populations.Enhancing closed-loop control: Enhanced closed-loop control refers to optimizing the communication between the user and the device, allowing for more natural and intuitive movements. This can be achieved through advanced sensors, machine learning algorithms, and real-time feedback. By improving the accuracy and response speed of the device, patients may experience greater control and improved function.Integrating multiple modalities: Furthermore, there is a growing need to explore the use of various modalities in virtual rehabilitation. Advances in this direction can result in the development of personalized multimodal interventions tailored to individual needs and preferences. Such interventions can integrate visual, auditory, and haptic feedback to create rich and immersive training experiences.Supporting reintegration: Reintegration support involves providing patients with the necessary resources and support to successfully reintegrate into society following amputation and rehabilitation. This may include vocational training, peer support groups, and mental health services. By addressing the psychosocial aspects of rehabilitation, patients may enjoy more fulfilling lives.Persistent attention has been devoted to serious games within the realm of rehabilitative practices over an extended period. Our study delves into the utilization of game-based approaches in the context of prosthetic rehabilitation with a specific emphasis on upper limb training. To ascertain the effectiveness of games in upper limb prosthetic rehabilitation, it is imperative to prioritize extensive, long-term experiments conducted within individuals’ homes, as they yield substantial evidence of their efficacy.Virtual environment paradigms should consider limb dominance to optimize their settings for better performance and perceptional engagement. Adapting rehabilitation for handedness may benefit unilateral impairments .Addressing these challenges will be critical to successfully translating virtual rehabilitation technologies into clinical practice, where they could significantly contribute to the goal of improving healthcare delivery for amputees. Moreover, the implementation of virtual prosthetic training in the field of prosthesis rehabilitation has primarily been restricted to academic research, resulting in a dearth of practical applications in clinical settings. This phenomenon is readily noticeable as there exists a gap in the translation of research outcomes to real-world scenarios. In addition, the efficacy of virtual technology in long-term rehabilitation following upper limb amputation calls for additional empirical evidence. Although virtual technology has shown significant benefits, establishing a standard protocol for virtual technology-based rehabilitation training is still very important to clinical use. Nevertheless, it can be foreseen that this technology will have great application prospects in various stages of the rehabilitation of upper limb amputees. The lack of a widely accepted framework for virtual prosthetic training could be attributed to the current limitations in data collection, data processing, and user interfaces. Implementing adequate protocols for the aforementioned challenges can ensure a smooth transfer of virtual prosthetic training techniques from academic research to clinical practice.Virtual environments are effective for prosthetic rehabilitation in amputees as they can accomplish control training through virtual environments and transfer their control experience to the real prostheses.The technological components outlined in this study, such as signal control, sensation feedback, adaptability, and gamification, can be combined for the purpose of enhancing user engagement and facilitating autonomous learning within and beyond clinical settings.The integration of immersive serious gaming and visual and somatosensory feedback using virtual technology, in particular, holds immense potential for promoting prosthetic embodiment within the virtual space.As discussed in this review, we have drawn the following conclusions:Virtual technology provides a flexible and configurable platform for upper limb amputees in their control rehabilitation in both admissions to the hospital and discharge. The virtual environment integrates multidisciplinary approaches associated with the timely and latest technologies from the fields of biomedical engineering and computer science. Long-term rehabilitation after discharge can also benefit from virtual restoration in terms of prosthetic adaptation, occupational rehabilitation, dignity recovery, and the reintegration of psychosocial status.


Estimations show that at least one in every 3 people in the world needs rehabilitation at some point in the course of their illness or injury. Access to rehabilitation services is an essential part of the continuum of care and is integral to achieving universal health coverage. However, most of the world’s population living in low- and middle-income countries, especially in the sub-Saharan African region, does not have access to adequate rehabilitation services. Wider adoption of digital solutions offers opportunities to support and enhance access to rehabilitation services in sub-Saharan Africa. A region where there is a greater burden and need for these services. There is also little published research about digital rehabilitation in sub-Saharan Africa, as it is an underexamined topic in the region.This scoping review aims to provide a comprehensive picture of the current evidence of digital interventions in rehabilitation implemented in any health, social, educational, or community setting in the sub-Saharan Africa region.We will conduct a scoping review using Arksey and O’Malley’s methodological framework and follow the Joanna Briggs Institute methodology for scoping reviews. We will develop search strategies for a selected number of web-based databases, search for peer-reviewed scientific publications until September 2023, and screen the reference lists of relevant articles. We will include research articles if they describe or report the use of digital interventions in the rehabilitation of patients with any health problem or disability in sub-Saharan Africa. For selected articles, we will extract data using a customized data extraction form and use thematic analysis to compare the findings across studies.The preliminary database search in MEDLINE (EBSCO) was completed in May 2023. The research team will conduct a search of relevant articles in the autumn. The results will be synthesized and reported under the key conceptual categories of this review, and we expect the final scoping review to be ready for submission in early 2024.We expect to find gaps in the research and a lack of detailed information about digital rehabilitation interventions in sub-Saharan Africa, as well as potential areas for further study. We will identify opportunities to inform the development of digital rehabilitation interventions.PRR1-10.2196/48952 Anyone with a health condition who experiences difficulties in any area of their life might need rehabilitation. According to the World Health Organization (WHO) , rehabilIt is estimated that at least one in every 3 people in the world needs rehabilitation at some point in the course of their illness or injury . AccordiRehabilitation has been effective in improving functioning outcomes in different health conditions in low-income and lower-middle-income countries, but it has not been prioritized in some of these countries and is underresourced . StrengtRehabilitation targets an individual’s needs and priorities in functioning across the continuum of care and throughout the individual’s life span. As countries move toward integrated person-centered care, it is imperative that quality rehabilitation be embedded in service delivery models . The comDigital technologies are considered an essential solution to health services in countries with limited resources around the world, including those in sub-Saharan Africa ,14. Sub-In the global context, rehabilitation models are costly and based on professionally delivered services. Most LMICs do not have a sufficiently educated rehabilitation workforce, leading to a situation where services do not meet demand . DigitalWe consider digital rehabilitation as an umbrella term, including different digital technologies that are used by rehabilitation professionals or self-driven service users as part of the rehabilitation process in its different phases . These include, but are not limited to, the use of tele- and distance rehabilitation applications and services  and the A multidisciplinary approach considers the person and their entire needs. Rehabilitation is simply not a health issue but includes education, social, livelihood, and empowerment-related efforts . If consWider adoption of digital solutions has the potential to improve access to rehabilitation services and avail more research opportunities, especially in LMICs where there is a greater burden and need for rehabilitation services ,23. SomeWe conducted a preliminary search of MEDLINE, the Cochrane Database of Systematic Reviews, and JBI Evidence Synthesis and did not identify current or ongoing systematic or scoping reviews on the topic. Unlike Kipruto et al’s  ongoing Therefore, the objectives of this scoping review are to identify the scope and types of evidence of digital rehabilitation solutions and to examine how research about digitally delivered rehabilitation interventions is conducted in sub-Saharan Africa. We expect the review to summarize the available evidence of digital rehabilitation interventions and identify potential research gaps and avenues for development. We chose a scoping review to explore the extent of literature on the topic and as a necessary step in identifying how digital rehabilitation technologies can be integrated into primary health care.We will conduct this review according to Arksey and O’Malley’s  frameworThe main research question guiding this review is: “What is the scope of research related to digital interventions used for rehabilitation in sub-Saharan Africa, and how is the research conducted?”To select relevant studies, we have developed eligibility criteria . These cInclusion criteriaPopulation: health professionals and patientsSocial, health, and other professionals delivering rehabilitation servicesPersons using or in need of rehabilitation servicesConcept: use, experiences, and issues of integrating digital services in rehabilitationStudies describing the use of digital interventions in rehabilitationStudies evaluating the effectiveness of various types of digital rehabilitation interventionsStudies reporting the problems and barriers to integrating digital services into rehabilitationContext: digital rehabilitation interventions adapted, piloted, and deployed in sub-Saharan AfricaStudies conducted in health, social, educational, and community settings in sub-Saharan AfricaTypes of sourcesPeer-reviewed articles, including research protocolsOther considerationsPublished in EnglishFull article is available on the internetExclusion criteriaConceptStudies reporting digitalization of health services other than rehabilitationTypes of sourcesEditorialsOpinion and perspective articlesAbstracts onlyLiterature reviewsThe review will include studies involving patients of all ages receiving treatment through digital rehabilitation interventions delivered by multidisciplinary rehabilitation professionals.The concepts of interest in this review are the use and effectiveness of digital rehabilitation interventions and how these interventions are implemented and evaluated in research. For the purposes of the review, we will only include digital rehabilitation interventions, such as telerehabilitation, wearable devices, robotics, virtual and augmented reality, smart device apps, and gamified mobile and web-based interventions.Studies conducted within any rehabilitation setting in sub-Saharan Africa will be eligible for inclusion.The search strategy will be limited to peer-reviewed literature. Primary research studies of all designs, including both quantitative and qualitative, will be included in this review. Secondary research studies  will not be included.We planned a search strategy to identify a comprehensive list of research literature specific to digital solutions used for rehabilitation in sub-Saharan Africa. Text in the title and abstracts of relevant articles, and the index terms used to describe the articles, were used to develop a full search strategy for MEDLINE EBSCO; . We willWe will collate and upload identified citations from all databases into Covidence   and remoThe research team will develop a data extraction tool to record the key information from the studies. Data will be extracted from papers included in the scoping review by 2 or more independent reviewers using the data extraction tool. The data extracted will include specific details about the participants, concept, context, study methods, and key findings relevant to the review question. These include author, publication year, publication type, country of origin, aims, population sample size and characteristics, methodology or methods, intervention type, outcomes and details of these, and other key findings that relate to the review’s question as suggested by the Joanna Briggs Institute . A draftThe research team will review the data extraction form and pretest it before implementation to ensure it is capturing relevant information. We will make modifications to the form after testing if necessary and state these modifications in the review. The extraction strategy will also be updated according to the categories emerging as the review progresses. We will resolve any disagreements that arise between reviewers through discussion and consensus, or with additional reviewers. The authors of the original papers will be contacted to request missing or additional data when required.Name of author, date of publicationAims of the study and research questionPopulationCharacteristics of participantsSample sizeCountry of origin (geography of the study)MethodologyStudy designRecruitment settingIntervention type, comparator, and details of these .Sampling strategy (if applicable)Key findings relating to the research question.ConclusionsWe will summarize the results using descriptive statistics. Descriptive statistics will be used to report the number of studies under each general category and additional conceptual categories, such as the study design, type of intervention, and the challenges and barriers to integrating digital solutions.A narrative summary will accompany the tabulated and charted results and will describe how the results relate to the review’s objectives and questions. Thematic analysis conducted according to Braun and Clarke’s  outline We will collect all data through searches of web-based scientific databases. No information on individuals will be collected, so approval from a research ethics committee is not required. The study follows the ethical principles of Jamk University of Applied Sciences.The protocol was registered and submitted for publication in May 2023. Databases will be searched in September 2023. Based on the preliminary search , we expeThe review will examine published peer-reviewed research about digital solutions for rehabilitation to establish the extent of their use. This will provide a comprehensive picture of the current knowledge on the use of digital rehabilitation interventions in sub-Saharan Africa and the potential for these interventions to improve patient outcomes. The review extends previous research by Kipruto et al  and NizeThis work is the initial step in a research project aiming to develop evidence-based, person-centered, and context-specific digital rehabilitation services in LMICs in East Africa and Southeast Asia. In the project, we aim to propose a digital-first multidisciplinary approach to address the challenges of access to rehabilitation services in low-resource settings. A digital-first approach will assist in delivering services directly to patients or through the primary health care system instead of solely through rehabilitation professionals, as is the case with the practice in high-income countries.A potential limitation is the availability of full-text articles. Some relevant articles may not yet be accessible during the search process; thus, limiting the reviewers’ ability to give a summary that reflects all available evidence of digital rehabilitation. Lack of relevant studies because of the sole focus on peer-reviewed research is another limitation. As the digital landscape goes through rapid changes, there may be limited information about studies related to digital rehabilitation interventions in the sub-Saharan context if they, for example, do not progress beyond the trial phase of a project after deployment. Focusing solely on English-only articles also limits the review, as there may be relevant publications in other languages spoken in the sub-Saharan African region, such as French or Portuguese.We expect the review to give us insights to develop the digital-first rehabilitation model proposed in the project. In addition, we expect it to outline research gaps and the implications for future research, policy, and practice. Other researchers, health professionals, and policy makers will benefit from the results. We will disseminate our findings through different channels, including peer-reviewed publications, presentations at scientific conferences, and web-based blogs.


Background: Augmented reality (AR) allows the overlapping and integration of virtual information with the real environment. The camera of the AR device reads the object and integrates the virtual data. It has been widely applied to medical and surgical sciences in recent years and has the potential to enhance intraoperative navigation. Materials and methods: In this study, the authors aim to assess the accuracy of AR guidance when using the commercial HoloLens 2 head-mounted display (HMD) in pediatric craniofacial surgery. The Authors selected fronto-orbital remodeling (FOR) as the procedure to test . Six people (three surgeons and three engineers) were recruited to perform the osteotomies on a 3D printed stereolithographic model under the guidance of AR. By means of calibrated CAD/CAM cutting guides with different grooves, the authors measured the accuracy of the osteotomies that were performed. We tested accuracy levels of ±1.5 mm, ±1 mm, and ±0.5 mm. Results: With the HoloLens 2, the majority of the individuals involved were able to successfully trace the trajectories of the frontal and nasal osteotomies with an accuracy level of ±1.5 mm. Additionally, 80% were able to achieve an accuracy level of ±1 mm when performing a nasal osteotomy, and 52% were able to achieve an accuracy level of ±1 mm when performing a frontal osteotomy, while 61% were able to achieve an accuracy level of ±0.5 mm when performing a nasal osteotomy, and 33% were able to achieve an accuracy level of ±0.5 mm when performing a frontal osteotomy. Conclusions: despite this being an in vitro study, the authors reported encouraging results for the prospective use of AR on actual patients. Augmented reality (AR) is a computerized technology which transfers virtual information to real environments. The technology is based on the “onlay” principle, i.e., the camera “reads” the object in the frame and the system recognizes it and activates a new level of communication, overlapping and integrating the virtual information with the actual object or environment . Unlike In recent years, its use in surgical navigation has been widely validated; many procedures, such as ventriculo-peritoneal shunt insertion and tumor resection in craniofacial and neurosurgery, are now usually performed using AR navigation .Standard surgical navigation relies on an external device that recognizes the patient’s position in the environment and then merges information from patient imaging, e.g., computer tomography (CT) or magnetic resonance imaging (MRI) on the screen.The navigator ensures that the noble anatomical structures can be localized before and during the operation. This improves the safety of the procedure. By means of navigation, it is also possible to import onto the patient’s imaging the trajectories of osteotomies and the target lesions in order to improve the extension and invasiveness of the surgery itself ,17.Though conventional navigation is a well-established methodology in surgery, it is burdened by several drawbacks. One of these is the fact that the operator has to switch his or her attention continuously between the patient and the screen ,12,15.AR head-mounted displays (HMDs) represent a technology that overcomes this limitation and can improve surgical navigation. Indeed, HMDs have integrated displays in order to allow surgeons to receive pertinent information while focusing their view on the surgical field . HMDs enCraniofacial surgery is a subspecialty that aims to address congenital skull dysmorphologies in pediatric patients. Among these, we cite single-suture craniosynostosis, multiple-suture synostosis, and syndromic craniosynostosis.It is a surgery that demands a certain amount of accuracy to avoid noble structures. Additionally, the morphological surgical outcome relies on the accurate design of the osteotomy .In this study, the authors aim to assess the accuracy of AR guidance when using the commercial HoloLens 2 HMD  in pediaIn particular, we focus on the procedure used to address frontal skull anomalies, known as fronto-orbital remodeling (FOR).While this is a pre-clinical study on a 3D printed phantom, the authors ultimately want to demonstrate the extent to which AR guidance using the HoloLens 2 smart glasses can accurately reproduce osteotomy trajectories in pediatric craniofacial surgery.This study was designed in order first to implement the AR-based protocol using the HoloLens 2 smart glasses. The authors then arranged a test session to evaluate the success rate in executing the AR-guided osteotomies for fronto-orbital remodeling (FOR) on a 3D printed phantom.In the following sections, the development phase and the experimental phase of the study are discussed.We selected the preoperative CT scan of a patient who had already been admitted and undergone an operation (study protocol CE 499-2022-OSS-AUSLBO). A DICOM file dataset was acquired and segmented in order to reconstruct a three-dimensional (3D) virtual model of the skull. Areas of the subject’s head that were of anatomical interest  were segmented using Mimics . Next, 3D meshes were generated from all the segmented masks and saved in standard tessellation language (STL) format .An appropriate portion of the reconstructed skull was selected for printing. In particular, relying on clinical information, we decided to visualize the skull from the top, as it would be viewed in theatre, and to cut the model behind the coronal sutures and bilaterally at the level of the fronto-zygomatic sutures.From the cut STL files, a phantom model made of photosensitive resin was produced by means of a stereolithography (SLA) 3D printer .To evaluate the AR guidance accuracy, CAD/CAM templates were designed using MeshMixer 3.5 software , and these were to be positioned on the surface of the phantom model, as cutting guides, in correspondence with the planned FOR osteotomies. For the AR-guided task, we selected the nasal and the frontal osteotomies that are part of the fronto orbital remodeling , left.The templates were 3D printed  with grooves of different widths  in order to evaluate three levels of achievable accuracy  , right. The virtual skull model with all its components , was imported into the Unity 3D software 2019.4.21f1 , extended with a specific software development kit for creating augmented reality apps .By means of the Vuforia Engine software, the registration between the virtual osteotomy traces and the skull phantom was achieved using the “model target” function, which allows the system to recognize the shape of an actual object to be tracked. In order to achieve this, the object has to be observed from a certain perspective by the surgeon wearing the AR glasses. In this case, we decided to reproduce the point of view of the surgeon in the theatre when performing the surgical procedure for fronto-orbital remodeling. The application draws and projects in front of the user wearing the AR glasses a profile (“guide view”) of the model target; the user simply needs to move the lenses and match the projected drawing on the actual object.In this study, a 3D model of the patient’s skull was used as the model target for the virtual-to-real scene registration.The AR application generates several holograms, which are superimposed on the printed skull portion, for each structure we want to visualize, i.e., the bony skull, the skin, the brain, the eye globes, and the FOR osteotomy trajectories to be used as guiding information during the surgical task.The AR application was built as a UWP  app deployed on the Microsoft HoloLens 2 smart glasses.Interactive user interface toggles (check boxes) were added to turn the rendering of each virtual anatomical structure and the planned virtual osteotomy trajectories on and off. Voice commands to show/hide the virtual structures were also implemented in order to provide a completely hands-free AR guidance system.The authors tested the AR application for the HoloLens 2 by having the selected FOR osteotomies, i.e., the nasal osteotomy (27 mm long) and the frontal osteotomy (75 mm long) performed under its guidance.We recruited three surgeons and three engineers . Each user repeated the procedure six times on the same 3D printed phantom with intervals of one week between each trial. Each user was briefed about the task that they were to perform. They had to perform the osteotomies on the phantom under the AR guidance provided by the HoloLens 2. Each user, after having calibrated the HMD for the perception of the optimal holograms, looked at the phantom in order to track it via the model target tracking function and then began the AR-guided task. Each user carefully drew the trajectory of the osteotomy on the skull phantom with a pencil, following the planned trajectories which were displayed as holograms in dashed lines. Vocal commands allowed the users to show/hide the virtual structures during the execution of their tasks.Afterward, using the 3D printed templates for accuracy evaluation, another operator assessed the extent to which the line traced under AR guidance fell into the groove of the single template. Each template had a calibrated tape along the groove itself to facilitate the measurements.All measurements were recorded in an Excel spreadsheet file. Percentages were recorded and both a Kruskal–Wallis test and a Mann–Whitney test were performed on the measurements.p value of <0.05 was considered statistically significant. SPSS software  was used to perform the statistical analysis, and a The results are summarized in For accuracy levels of ±1 mm and ±0.5 mm, lower success rates were recorded. Specifically, for the nasal cut, we reported success rates of 80% and 61%, respectively. Regarding the frontal cut, the users were able to accomplish the task with an accuracy level ± 1 mm in 52% of cases, whereas only 33% precisely followed the groove with an accuracy level of ±0.5 mm.The Kruskal–Wallis test demonstrated that all the users were able to complete the nasal cut task with no significant differences between them. However, more inter-operator differences were reported for the frontal cut .From the measurements, only one outlier was evident according to the Mann–Whitney test .The users reported that the usability of the AR guidance system was very good, but most of them reported a perceived loss of image quality when moving the pencil in front of the visor.AR is a promising technology in the medical field; more and more studies on its applications, especially in the surgical field, are being published due to increasing interest ,27,28,29Its introduction enables the realization of a full concept of navigation, i.e., the operator does not have to shift his or her attention anymore and can stay focused on the patient, on whom holograms are projected. A specific taxonomy was introduced in the 1960s to categorize this technology .AR HMDs are divided into optical see-through devices and video see-through devices ,29,30,31Craniofacial surgery, and in particular corrective procedures for forehead morphological anomalies, can be challenging due to the noble structures underlying the bone . FurtherTherefore, an HMD suitable for this surgery has to be accurate and maintain the hologram in the field of vision even if the surgeon moves. The HoloLens has already demonstrated good potential in this sense ,24.However, different drawbacks and limitations have been reported in the literature, such as depth of perception and registration errors ,32.We have reported some drawbacks, too. In an effort to overcome these limitations, our group has already tried to address the registration errors, in which a static error ranging from 1 mm to 10 mm is reported. This results in a misalignment in the subjective perception of the virtual image and actual image . This reIn this study, the authors evaluated the accuracy of the HoloLens 2 when used to perform a craniofacial surgery task. The procedure of choice was fronto-orbital remodeling, focusing in particular on two osteotomies, nasal and frontal, which define the orbital rim.The authors selected six operators, and every operator had to repeat the task six times for each osteotomy, both nasal and frontal. In order to avoid an additive learning curve effect, an interval of one week passed between each trial.The users were asked to trace the osteotomies with a pen under the guidance of the HoloLens 2 projection. They could see on the lenses the phantom and the dotted lines for the planned osteotomies. We noticed some errors in virtual-to-real alignment and a loss of sharpness when moving the pen in the field of view.All the users were able to complete the task. The osteotomy traced with the pen was then checked with the cutting guides having different width grooves.In terms of accuracy, our findings are consistent with previous findings outlined in the literature, and, to a certain extent, this is encouraging. We described the maximum accuracy with an error of ±1.5 mm. We also noticed that a lower level of accuracy was reported for the frontal cut, and this might be due to the length of the trace that had to be followed (75 mm), and to the more complex round anatomy.Despite these technical pitfalls, the levels of accuracy that we reported are consistent with what has been described elsewhere in the literature ,33,34,35Scherl et al. reported an accuracy of less than 1.3 mm in their in vivo study , whereasHan et al. described their experience in craniosynostosis, and this is the only available on-patient work of its kind in the literature. Their study involved seven patients undergoing calvarial remodeling for plagiocephaly, and they compared the planned intracranial volume with the obtained one, with encouraging results . In our Our findings have, however, been encouraging. Our results included only one outlier, and therefore we cannot exclude a learning curve effect.Our study had limitations, principally due to the advantageous lighting conditions compared with those typically found in theatre, the errors in model target registration, and operator-dependent factors. The authors also wish to emphasize that, despite the fact that loupe glasses are in common use, HMD technology has not yet been optimized for compatibility with loupes. Furthermore, the procedures were performed in ideal conditions. Our next step will be to implement the navigation system by inserting more details and more 3D objects, to be seen contemporaneously with the trajectories that are to be followed with the instruments . The preparation time for the 3D model reconstruction, starting from the DICOM data segmentation and including the setting up of the AR guidance software, was only one to two hours . An in vThis was an in vitro study. The encouraging results in terms of the ±1.5 mm accuracy make it suitable for application in craniofacial surgery, where such a margin of error can be admitted in certain tasks. More studies, including in vivo evaluations, are required to overcome the technical pitfalls involved in this promising technology.


Mechanistically, lnc-AROD interacted with miR-324-5p, leading to decreased binding of miR-324-5p to CUEDC2. Collectively, our findings demonstrated that lnc-AROD is a critical regulator of the host antiviral response via the miR-324-5p–CUEDC2 axis, and lnc-AROD functions as competing endogenous RNA. Our results also provided evidence that lnc-AROD serves as an inhibitor of the antiviral immune response and may represent a potential drug target.Long noncoding RNAs (lncRNAs) are a class of noncoding RNAs that are involved in multiple biological processes. Here, we report a mechanism through which the lnc-AROD–miR-324-5p–CUEDC2 axis regulates the host innate immune response, using influenza A virus (IAV) as a model. We identified that host lnc-AROD without protein-coding capability is composed of 975 nucleotides. Moreover, lnc-AROD inhibited interferon-β expression, as well as interferon-stimulated genes ISG15 and MxA. Furthermore, IMPORTANCE lnc-AROD is a potential diagnostic and discriminative biomarker for different cancers. However, so far the mechanisms of lnc-AROD regulating virus replication are not well understood. In this study, we identified that lnc-AROD is downregulated during RNA virus infection. We demonstrated that lnc-AROD enhanced CUEDC2 expression, which in turn inhibited innate immunity and favored IAV replication. Our studies indicated that lnc-AROD functions as a competing endogenous RNA that binds miR-324-5p and reduces its inhibitory effect on CUEDC2. Taken together, our findings reveal that lnc-AROD plays an important role during the host antiviral immune response. Because most studies have focused on the function of proteins, less is known about the role of RNA, including noncoding RNA (ncRNA) . High-th17–The lncRNA-activating regulator of Dickkopf WNT signaling pathway inhibitor 1 (DDK1) lncRNA (lnc-AROD), a spliced and polyadenylated lncRNA, has been reported to positively regulate DKK1 transcription at the level of transcription elongation, as well as recruit the general transcription factor EBP1 to the DKK1 promoter . Some st23–Recently, lncRNAs have been found to be involved in modulating viral infection and the innate immune response . lncRNA-In the present study, genome-wide profiling of lncRNA expression identified an IAV-dysregulated novel lncRNA, termed lnc-AROD, which was capable of favoring IAV replication. lnc-AROD was found to function as a negative regulator in host antiviral immunity by suppressing ISG15 and MxA production. Further data demonstrated that lnc-AROD acts as a competing endogenous RNA (ceRNA) to enhance the level of CUEDC2 expression through sponging with miR-324-5p. Our results revealed that lnc-AROD is a negative regulator of IFN-β and ISG expression, establishing a critical role for this lnc-RNA in the host innate defense during IAV infection.High-throughput RNA sequencing (RNA-seq) was performed to determine the lncRNA expression profiles in H1N1 virus-infected A549 cells at 18 h postinfection (hpi). Based on the specific location in the genome, the inclusion of multiple exons, and a length greater than 200 nucleotides (nt), noncoding transcripts were filtered to identify the novel lncRNAs. Moreover, noncoding transcripts were classified by CPAT, PFAM, and CPC. As a result, 258 distinct lncRNA candidates were selected  and B. FTo explore crucial lncRNAs involved in a viral infection, hierarchical clustering was performed to determine the lncRNA expression profiles in A549 cells infected with or without the H1N1 virus for 18 h. The level of lncRNA expression was significantly altered following H1N1 infection . The dif50) assays revealed that the virus titers from lnc-AROD-A549 cells were significantly higher than those in the control cells  expressing lnc-AROD or green fluorescent protein (GFP) were generated to express human lnc-AROD in mice. AAV–lnc-AROD and AAV-GFP (control) were intranasally administered to mice for 21 days, followed by the delivery of phosphate-buffered saline (PBS) or H1N1 virus (103.5 TCID50) on day 0  assay also demonstrated that lnc-AROD interacted with miR-324-5p . Thus, we selected the predicted CUEDC2 as a potential target gene of miR-324-5p on TargetScan and miRanda. To determine whether the lnc-AROD–miR-324-5p axis involves CUEDC2 during IAV replication, we examined CUEDC2 expression after virus infection. The results from the qPCR assay showed that IAV infection could stimulate CUEDC2 downregulation . The seq50 assays. The data from The above data demonstrated that miRNA-324-5p directly interacts with lnc-AROD and CUEC2. Considering that lncRNA combines with miRNA as a ceRNA , 39, we Consistently, the viral titer was significantly decreased in lnc-AROD-A549 cells with CUEDC2-specific siRNA transfection but not in CUEDC2-overexpressing cells after H1N1 infection, compared to that of control siRNA  and J. Min vitro and in vivo lnc-AROD overexpression, but not knockdown, facilitated RNA virus replication and not DNA virus replication. Notably, lnc-AROD overexpression significantly increased the mortality rate in mice during RNA virus infection. These findings suggested that lnc-AROD might be important for IAV pathogenesis in vivo and that it functions as a key factor via negatively regulating the expression of some crucial antiviral proteins.lncRNA expression profiles have been found to display a large amount of lncRNA data. ln-AROD is upregulated in different tumor tissues and cells –25, and –miR-324-5p plays an important role in cancer, disease, and differentiation by interacting with noncoding RNA and proteins. miR-324-5p regulates mitochondrial morphology and cardiomyocyte cell death by targeting Mtfr1 . MoreoveCUEDC2, a CUE-domain-containing protein that modulates inflammation, cell cycle, and tumorigenesis, can negatively regulate the JAK-STAT pathway to induce type I and type III interferons . In the lnc-AROD, CUEDC2, and lnc-AROD-ORF1/ORF2-FLAG were cloned into the pCDH-CMV-MCS-EF1-copGFP plasmid ; these constructs were named pCDH-AROD, pCDH-ORF1, pCDH-ORF2, and pCDH-CUEDC2. The full-length lnc-AROD, lnc-AROD-del, CUEDC2–3′-UTR, and CUEDC2–3′-UTR-del were inserted into the pGLO plasmid  and were named pGLO-AROD, pGLO-AROD-DEL, pGLO-CUEDC2–3′-UTR, and pGLO-CUEDC2–3′-UTR-del. Argonaute2 (AGO2) was cloned into the PCDH-3flag-EGFP plasmid , and lnc-AROD was cloned into the pMS2 plasmid, which was gifted from Myriam Gorospe . lnc-ARO2 atmosphere.Human embryonic kidney HEK293T (293T) cells and A549 cells were obtained from the Shanghai Institute of Cell Biology, Chinese Academy of Sciences . lnc-AROD-overexpressing 293T and A549 cell lines were produced by cotransfection with pCDH-AROD, the helper lentiviral packaging plasmids psPAX2.0  and pMD2.G . All cells were cultured in Dulbecco’s modified Eagle’s medium  supplemented with 10% fetal bovine serum . Cells were incubated at 37°C in a humidified 5% COInfluenza virus A/Puerto Rico/8/34 (H1N1), A/Quail/Hangzhou/1/2013 (H9N2), A/Swine/Zhejiang/04/2009 (H3N2), A/Hangzhou/1/2013 (H7N9), A/pika/Qinghai/BI/2007 (H5N1), and VSV were stored by our laboratory and propagated in 9-day-old embryonated eggs at 37°C. SeV and HSV were propagated in 293T cells.A549 cells were infected with H1N1 at a multiplicity of infection (MOI) of 10 for 18 h. The total RNA was extracted by TRIzol reagent  according to the manufacturer’s instructions. rRNAs were further removed using a ribo-zero-magnetic-kit . RNA-seq libraries were prepared with a TruSeq RNA LT sample prep kit v2  and sequenced on the Illumina Hiseq 3000 platform  at Shanghai Genergy Biotech . Samples were divided into two groups: H1N1-infected and uninfected (mock) cells. Each group contained three biological duplicates.http://www.bioinformatics.babraham.ac.uk/projects/). Fast QC_v0.11.5 software (http://www.bioinformatics.babraham.ac.uk/projects/fastqc/) was used for a quality control analysis. The remaining reads were mapped to a human reference genome (version Homo_sapiens GRCh38) with the STAR_v2.5.2b software suite (http://pfam.xfam.org/), coding potential calculator  were filtered out by Trim Galore_v0.4.2 . The cDNA was synthesized using a Thermo Scientific RevertAid first-strand cDNA synthesis kit  to quantify the amount of mRNA and lnc-AROD. Glyceraldehyde 3-phosphate dehydrogenase (GAPDH) was used as an internal control. The cDNA was synthesized using a miRNA first-strand cDNA synthesis kit  to quantify the amount of miRNA. Small nuclear U6 RNA was used as an internal standard. Real-time PCR analyses were performed using ChamQ Universal SYBR qPCR master mix . To determine the absolute quantity of RNA, the purified PCR product amplified from the cDNA corresponding to the lnc-AROD and IAV-M sequences was serially diluted to generate a standard curve. The primers used in this study are listed in Table S2.The 5′ and 3′ RACE procedures were performed using a SMARTer RACE cDNA amplification kit  in accordance with the manufacturer’s protocol. The primers for lnc-AROD are listed in Table S1. The RACE PCR products were inserted into the pMD 18-T simple vector  and sequenced.Renilla luciferase activity) was calculated.The 293T cells were seeded into 24-well plates and transfected with pGLO-AROD, pGLO-AROD-DEL, pGLO-CUEDC2–3′-UTR, or pGLO-CUEDC2–3′-UTR-DEL together with miR-324-5p mimics. After 48 h, the luciferase activity was measured with a dual luciferase reporter gene assay kit  according to the manufacturer’s protocol. Relative luciferase activity  supplemented with RNase  and protease inhibitors for 15 min. The lysates were incubated with magnetic beads conjugated with IgG (as a control) and an anti-Flag antibody  at 4°C for 4 h. The beads were washed five times and lysed in TRIzol reagent for RNA extraction and subjected to RT-qPCR to determine the amount of lnc-AROD.2, 0.05% NP-40, 1 mM phenylmethylsulfonyl fluoride, 10 mM ribonucleoside vanadyl complex) supplemented with RNase  and protease inhibitors for 15 min. The lysates were incubated with GST-magnetic beads at 4°C for 4 h and washed five times, and RNA was extracted from the beads followed by RT-qPCR to quantify the amount of miR-324-5p.An MS2-RIP assay was performed as previously reported . Brieflylnc-AROD-overexpressing 293T cells were seeded into six-well plates in three replicates and transfected with the 3′ end of biotinylated miR-324-5p mimic or control RNA  for 24 h, as previously described . BrieflyThe specific probe for the lnc-AROD sequence was synthesized by GenePharma , and miR-324-5p was transcribed to RNA fragments with a TranscriptAid T7 high-yield transcription kit (Thermo Scientific) and labeled with Alexa Fluor 488 using a ULYSIS nucleic acid labeling kit .A549 cells were fixed with cold 4% paraformaldehyde for 30 min, incubated with 100% ethanol at 4°C for 12 h, and permeabilized with 0.5% Triton X-100 for 5 min. Wash buffer  was added and subsequently incubated with hybridization buffer  containing probe at 37°C for overnight. Nuclei were stained with 4′,6-diamidino-2-phenylindole (DAPI) and washed with PBS. Finally, images were obtained using a Zeiss LSM 880 laser confocal microscope. The probes used for FISH are listed in Table S1.Cells were washed twice with ice-cold PBS and lysed in lysis buffer . Samples were boiled and separated by 12% SDS-PAGE and transferred onto nitrocellulose membranes , followed by incubation with specific antibodies.n = 11); GFP–lnc-AROD + PBS group (n = 11); GFP + H1N1 group (n = 16); and GFP–lnc-AROD + H1N1 group (n = 16). To determine the level of lnc-AROD expression in the lungs, AAV-expressed lnc-AROD was produced by Vigene Biosciences, Inc. Briefly, lnc-AROD or negative sequences were inserted into an AAV6 vector, and the resultant vector was cotransfected with the helper packaging plasmids pAAV-RC and pHelper into 293T cells. After iodixanol gradient purification, the mice were intranasally administered approximately 25 μL (2.34 × 1013 v g/mL) AAV6-expressing lnc-AROD or negative control for 3 weeks. The mice were treated with 50 μL PBS or intranasally challenged with the H1N1 virus (103.5 TCID50) to establish a mouse model of IAV infection.Four-week-old male C57BL/6 mice were purchased from Shanghai SLAC Animal Company  and maintained in specific-pathogen-free barrier facilities. A total of 54 mice were randomly divided into 4 groups: GFP + PBS group  software. All data are reported as means ± standard deviations (SD). The differences between groups were analyzed with a Student's The animal experiment was approved by the Institutional Animal Care and Use Committee (IACUC) of Zhejiang University (SYXK 2012-0178). All animal experiments were conducted in accordance with the Regulations for the Administration of Affairs Concerning Experimental Animals approved by the State Council of the Peoples Republic of China.GSE208546).The RNA sequencing data were deposited in the Gene Expression Omnibus database (accession number


The past 2 decades have seen rapid development in the use of robots for rehabilitation. Research on rehabilitation robots involves interdisciplinary activities, making it a great challenge to obtain comprehensive insights in this research field.We performed a bibliometric study to understand the characteristics of research on rehabilitation robots and emerging trends in this field in the last 2 decades.Reports on the topic of rehabilitation robots published from January 1, 2001, to December 31, 2020, were retrieved from the Web of Science Core Collection on July 28, 2022. Document types were limited to “article” and “meeting” (excluding the “review” type), to ensure that our analysis of the evolution over time of this research had high validity. We used CiteSpace to conduct a co-occurrence and co-citation analysis and to visualize the characteristics of this research field and emerging trends. Landmark publications were identified using metrics such as betweenness centrality and burst strength.P<.001; tS=175.0) and citations  related to rehabilitation robots exhibited a significantly increasing yearly trend. The co-occurrence results revealed 120 categories connected with research on rehabilitation robots; we used these categories to determine research relationships. The co-citation results identified 169 co-citation clusters characterizing this research field and emerging trends in it. The most prominent label was “soft robotic technology” (the burst strength was 79.07), which has become a topic of great interest in rehabilitative recovery for both the upper and lower limbs. Additionally, task-oriented upper-limb training, control strategies for robot-assisted lower limb rehabilitation, and power in exoskeleton robots were topics of great interest in current research.Through data retrieval, cleaning, and deduplication, we retrieved 9287 publications and 110,619 references cited in these publications that were on the topic of rehabilitation robots and were published between 2001 and 2020. Results of the Mann-Kendall test indicated that the numbers of both publications (Our work provides insights into research on rehabilitation robots, including its characteristics and emerging trends during the last 2 decades, providing a comprehensive understanding of this research field. The past 2 decades have seen rapid, vast development of robots for rehabilitation. Rehabilitation robots are representative of advanced modern rehabilitation devices; they are automatically operated machines used to treat patients with impaired motor function -5. ReseaLiterature reviews provide researchers with a comprehensive understanding of particular areas of research . Most exRecently, improvements in computer and information science have enhanced bibliometric analysis and allowed an intensive interpretation of emerging trends in single and multiple research fields -21. The In this study, we performed a bibliometric analysis, including a co-occurrence and co-citation network analysis, of research conducted between 2001 and 2020 on robots for rehabilitation. Our aim was to understand developments in this research field and identify emerging trends.The bibliometric data used in this study were derived from scientific literature indexed by the Web of Science (WoS) Core Collection as of July 28, 2022. A comprehensive search strategy was used to meet the requirements for data coverage. This strategy involved both index terms and keywords, including truncation, proximity, and phrases. Terms such as “rehabilitation robot” were searched for as “rehabili* robot*” to identify all related terms. Document types were limited to “article” and “meeting,” excluding documents classified as “review” papers. The records included basic attributes of the documents, such as publication time, author, institution, country, and cited references; these were used to form a database that was used for the subsequent analysis.In this paper, CiteSpace, a Java application developed by Chen , was usetS. A positive tS value means that the trend is increasing, and a negative tS value means that the trend is decreasing.A Mann-Kendall test was used to assess whether the literature data, including publications and citations, increased year over year, and whether trends were statistically significant. For each comparison pair (publications or citations in 2 adjacent years), we assigned a score of +1 if the latter value was greater than the former value and a score of –1 if the latter value was lower than the former value. All scores were then summed to calculate the test statistic, A co-occurrence analysis was performed to observe the relationship between shared words in the literature. The frequency of word occurrence is associated with the underlying themes. In this analysis, a co-occurrence clustering network was generated to examine changes in specific topics in research on the use of robots in rehabilitation ,34. A coThe co-occurrence network and co-citation networks were mainly characterized by 2 metrics, betweenness centrality (BC) and burst strength (BS). BC was used to identify pathways between different thematic clusters. This computation was based on a fast algorithm introduced by Brandes  that allP<.001; tS=175.0) and citations  related to rehabilitation robots have exhibited a significantly increasing yearly trend in the last 2 decades. In the past 10 years (2011-2020), there is an obvious, fast-growing trend in the number of citations. The growth rate of citations is faster than that of publications.After data retrieval, cleaning, and deduplication, we retrieved a total of 9287 publications and 110,619 cited references in the field of research on robots for rehabilitation that were published from 2001 to 2020; we used these data to form a database . The numrobotics, (2) electrical and electronic engineering, (3) biomedical engineering, and (4) automationandcontrol system. The major categories in medical fields were rehabilitation, neurosciences, clinical neurology, sport sciences, and medical informatics. Each publication indexed in the WoS is assigned to one or more categories. Based on the classification of categories in the database , publicaBased on the literature data, a network of co-citation clusters was constructed and visualized  that consoft robotic) is currently a research hotspot, while cluster 1 (following stroke) and cluster 2 (robot-assistant gait training) have gradually lost attention. Recently, few reports have been published on topics related to the robotic treatment of patients with spinal cord injury or Parkinson disease.Further, we used the timeline view to visualize the evolution of the publications . In thisBC and BS are two important metrics in the co-citation analysis. In this study, we performed a bibliometric analysis of research on rehabilitation robotics published in the last 2 decades; we obtained insights on emerging trends in this research field. Our analysis indicates that the emergence of new technologies, such as virtual reality, brain-computer interfaces, and intelligent sensing, is advancing the development of rehabilitation robotics. Robots with a flexible structure are currently attracting great attention in robot design.Literature  retrieved from the WoS database between 2001 and 2020 shows a continuous increase in research interest in rehabilitation robotics. We excluded review papers from the literature data because they might have interfered with the bibliometric analysis; compared with original-research papers, review articles are more likely to be identified as prominent publications because they are usually highly cited by other researchers. However, most review papers are not associated with technical keywords. Even if some review papers provide technical keywords, they lag behind the publication date of the paper. Consequently, specific technologies identified as being important and having an increasing trend likely do not match the period of their actual emergence. For this reason, we excluded review papers from our document retrieval.Based on the literature data, we performed co-occurrence and co-citation analyses to characterize emerging trends in research on rehabilitation robots. The co-occurrence analysis of categories showed that a majority of articles on rehabilitation robots are published in fields related to engineering, such as robotics, electrical and electronic engineering, and biomedical engineering. The development of rehabilitation robotics has been going on for many years, and technological progress will continue in the coming decades. Newly emerging engineering technologies  are key drivers of advances in the development of rehabilitation robotics ,66,67. Fsoft robotic, hand rehabilitation, chronic stroke, exoskeleton robot, and gait rehabilitation. This cluster emerged between the years 2009 and 2019. Recently, soft robotics in rehabilitation have become a novel branch of robotics; they are characterized by compliant and flexible materials  [The co-citation analysis identified 169 clusters, allowing us to characterize and interpret the structure and dynamics of co-citation in research on rehabilitation robots. Specifically, we identified 7 prominent clusters that represent important research themes. The largest cluster  . Comparenylon 6) . Such flnylon 6)  that repnylon 6) . Anothernylon 6) . Althougnylon 6) . Additionylon 6) . Clusternylon 6) . Robot anylon 6) . ClusterLimitations of this bibliometric analysis include, first, that it was based on literature data retrieved only from the WoS database. This database might not include publications that were present in other databases, such as PubMed, Google Scholar, and Scopus, meaning that they were missing from our analysis. The main reason for selecting the WoS was that this database is designed to allow citation analysis and provides more extensive information ,78,79. RThis study identified prominent literature on the use of robots in rehabilitation through bibliometric analysis. We visualized and characterized co-occurrence and co-citation networks of publications in this research field, providing insights into the characteristics of the research and emerging trends over the last 2 decades. Our co-occurrence analysis showed that the emergence of new engineering technologies  advances the development of rehabilitation robotics. Our co-citation analysis indicates that flexible-structure robots are currently gaining wide attention for rehabilitative applications. Rehabilitation robots have been used most extensively in the treatment of chronic stroke. It is foreseeable that research on rehabilitation robots in the coming years will enjoy explosive growth that will promote extensive applications in improving motor function and quality of life in human patients.


However, when stratifying subjects by dairy SLPF intake, children in the highest tertile (>5%) showed a higher intake of sugars, total and saturated fats, and higher plasma levels of total and low-density lipoprotein cholesterol than those in the first tertile (<1%) (p < 0.05). The variability in the nutritional composition of SLPFs highlights the need for up-to-date databases which would greatly assist in optimizing individualized recommendations for children with IEMs and protein restrictions.In inborn errors of intermediate protein metabolism (IEM), the effect of special low-protein foods (SLPFs) on dietary intake has been scarcely studied. The aim of this study was to compare the nutritional profile of SLPFs with usual foods and to assess whether their intake determines the dietary pattern and affects the plasma biochemical profile in children with IEMs with different protein restrictions. A database with the nutritional composition of 250 SLPFs was created. A total of 59 children with IEMs were included in this cross-sectional observational study. The greatest significant differences in macronutrient composition were observed between dairy, meat, fish, and egg SLPFs and regular foods. After stratifying subjects by SLPFs, the participants with the highest intake (>32%) had a higher total energy intake and lower intake of natural protein than those in the lowest tertile (<24%) ( Disorders of protein and amino acid metabolism, such as phenylketonuria (PKU), maple syrup urine disease (MSUD), tyrosinaemia (TYR), homocystinuria (HCU), organic acidaemias (OAAs), and urea cycle disorders (UCD), are a group of inherited metabolic conditions caused by the deficiency of enzymes or transporters involved in amino acid metabolism. Clinical phenotypes range from asymptomatic to life-threatening . Due to In most of these pathologies, the primary treatment approach is dietary modification , in addiRegular monitoring of plasma amino acid levels is necessary to ensure that they remain within optimal ranges or to adjust vitamin or cofactor supplementation to increase enzyme activity when residual enzyme activity remains . In addin = 73), Belgium (n = 92), Italy (n = 256), and Germany (n = 94), while 146 products were reviewed in the United Kingdom [The dietary approach for managing these metabolic disorders typically involves calculating and measuring the natural protein source, adding carbohydrates and fats to the diet to increase caloric intake and prevent catabolism, and supplementing with calculated amounts of PFAAs as needed, along with SLPFs that provide energy and variety to the diet . SLPFs a Kingdom . However Kingdom . Althoug Kingdom , Metabol Kingdom , or the  Kingdom , no dataThe nutrients provided by each group of foods in a healthy diet are known; however, the contribution of different food groups and the consumption of SLPFs in the total diet of individuals with inborn errors of intermediary protein metabolism (IEM) with restricted protein diets are not clear. In the case of phenylketonuria, it has been estimated that the intake of SLPFs ranges between 30% and 60% of total calories ,11,12. HIt has been reviewed by Verduci et al.  that PKUTherefore, the aim of this study is to compare the nutritional profile of SLPFs with that of regular foods and to evaluate the extent to which the intake of SLPFs determines dietary patterns and plasma lipid and glucose profiles in a group of children with IEMs with different degrees of protein restriction.A total of 250 SLPFs were collected from Spanish manufacturers and suppliers, and an in-house database was created. Products were divided into 15 categories and included milk replacers; ice cream/whipped cream; cheese; breakfast cereals; bread and mixes; flour; pasta; rice; cookies; cakes and desserts; chocolate; ready meals; and meat, fish, and egg replacers.® [The nutritional information was collected from the mandatory nutritional labelling of the products or from technical data sheets when available. Nutritional data were obtained per 100 g or 100 mL of products for: energy; protein; total carbohydrate; sugars; fibre; total fat; saturated fatty acids (SFAs); and sodium or salt. Following the criteria of Wood et al. , when nu® , containThis was a cross-sectional observational study conducted from January 2021 to March 2022. Eligible patients for this study were children diagnosed with IEMs by newborn screening or genetically confirmed, treated with protein-restricted diets, aged 10 months to 17 years, without a language barrier or difficulty of understanding, and managed at Congenital Metabolic Disease Unit of Sant Joan de Déu Hospital (HSJD), as well as who continuously attended clinical appointments. In total, 59 children (27 girls and 32 boys) were recruited. Of these children, 30 had phenylketonuria (PKU); 9 had PKU tetrahydrobiopterin responders (PKU-BH4); 2 had maple syrup urine disease (MSUD); 4 had classical homocystinuria (HCU); 4 had organic acidaemias (OAAs) ; 1 glutaric aciduria type 1 (GA1) and 2 with 3-hydroxy-3-methylglutaric aciduria (3-HMG)); 4 had urea cycle disorders (UCDs) (which included 2 argininosuccinate synthetase deficiency (ASS), 1 with ornithine transcarbamoylase deficiency (OTC) and 1 with hyperornithinaemia, hyperammonaemia, and homocitrullinuria (HHH)); 3 had hereditary tyrosinaemia type I (HTI) and 3 had a liver transplant including 2 with deficiency of argininosuccinate lyase (ASL) and 1 with propionic acidaemia (PA).The study was approved by the Research Ethics Committee (CEIm) of the Fundació de Recerca Sant Joan de Déu (codeCEIm PIC-63-21). The parents or legal caregivers for all children provided their written informed consent before their inclusion in the study.https://www.seghnp.org/nutricional (accessed on 31 May 2022), based on the Barcelona Longitudinal Growth Study 1995–2017 [Weights and lengths for children under 2 years of age were obtained by standard techniques using digital baby-weighing scales and crown–heel length on a scaled length board. Heights and weights of children older than 2 years were measured using a combined stadiometer and a digital weight-measuring station . Height to the nearest 0.1 cm and weight to the nearest 0.1 kg were recorded. Anthropometric measurements were expressed as age- and sex-specific Z-scores, using the SEGHNP Paediatric Nutritional Application, available online: 995–2017 .Parents completed a 3-day food record (3DFR) of their children, including one nonworking day . The trained metabolic dietitian calculated calories and macro- and micronutrient intake of the 3DFR using the nutrient analysis software program ODIMET , which iStatistical analyses were performed using SPSS version 27.0 software . The Mann–Whitney non-parametric unpaired test was used to compare energy, nutrients , and sodium contents from SLPFs and regular foods. These comparisons were presented as a mean and standard deviation (SD).p-value < 0.05 was considered significant.The anthropometric parameters, dietary intake, and plasma biochemical data were skewed (Kolmogorov–Smirnov and Levene tests) and the natural logarithm of the variables did not normalize them; therefore, the median  was used to describe them. Spearman’s correlation was used to study the relationship between variables. Participants were also categorized based on tertiles of SLPF consumption and tertiles of dairy SLPF consumption. To study the differences across tertiles, data were analysed using the Kruskal–Wallis and Mann–Whitney post hoc tests or a Chi-square test. In addition, post hoc power analysis was used to calculate the statistical power of total cholesterol and LDL-C parameters according to tertiles of dairy SLPF consumption ,27. WhenFrom the nutritional analysis, the fibre values were obtained for all the groups except for dairy SLPFs due to unavailability of this parameter in technical data sheets. Some 60% of food groups showed significant differences in fibre. The most significant groups were meat, egg and fish replacers, which contained more fibre than regular foods  since regular products had no fibre content. In terms of sodium evaluation, 53% of the groups showed significant differences between regular foods and SLPFs. While significant higher levels of sodium were observed in meat replacers, low-protein pasta, flour and cheese, and a trend in fish replacers, there were other food SLPF groups—such as milk replacers, bread, cookies, and cakes—with significant lower amounts than regular foods.Therefore, globally, it has been observed that the SLPF groups of cheese, meat, fish, and egg replacers had higher differences in macronutrients with those of regular foods than the other food groups.n = 7), MSUD (n = 1), and a PA liver transplanted (n = 1); the five subjects who showed a BMI Z-score of between 1.5 and 1.95 were subjects with PKU (n = 1) and HTI (n = 1); the seven subjects who showed a BMI Z-score greater than 2 were subjects with PKU (n = 5), PKU-BH4 (n = 1), and MSUD (n = 1). On the other hand, one participant with an ASL liver transplanted showed a BMI Z-score of between −1 and −1.49; three participants showed a BMI Z-score of between −1.5 and −1.95, two participants with PKU and one with OAA; and two participants in total, one with PKU-BH4 and one with HCU, had a BMI Z-score lower than −2.0, indicating underweight.A total of 59 infants with a median age of 8.9 years were recruited. The median and IQR of plasma glucose, triglycerides (TAGs), total cholesterol (TC), high-density lipoprotein cholesterol (HDL-C), and low-density lipoprotein cholesterol (LDL-C) were within the normal range in the complete study population  considerThe results showed that 29% of the energy of the dietary intake occurred from the intake of SLPFs, while the 26% comes from the PFAAs intake, which both resulted in 55% of the energy of the total diet. Overall, and as expected, SLPF consumption made a very low contribution to protein intake compared to PFAAs, which provided the highest protein content for patients. It has to be emphasized that similar amounts of carbohydrates were provided by the SLPFs (21%) and from regular foods (22%) (dietary intake minus SLPFs minus PFAAs) . OtherwiIn this study, we mainly focused on dietary intake, and anthropometric and plasma biochemical parameters across tertiles of percentage of SLPF consumption . Particip < 0.05) and with total energy from PFAAs (p = 0.05). In addition, the consumption of SLPFs was positively correlated with the intake of protein from PFAAs. This could indicate that subjects that used more SLPFs could need the PFAAs in order to fulfil their protein requirements.However, significantly positive correlations were observed between participants who consumed more energy from SLPFs, and the ingested carbohydrates, sugars, and fats from them . As expep < 0.05), and total fats from SLPFs were positively correlated with carbohydrates, sugars, SFAs, MUFAs and PUFAs (p < 0.05).The relationship between carbohydrates, sugars, total fats, and SFAs derived from SLPFs and the other dietary components was also evaluated . It is tp = 0.04), total fats , and SFAs .However, in the first part of this work, it was concluded that SLPF dairy, meat, fish, and egg replacers had higher differences in macronutrients than the other food groups. In this regard, the consumption of dairy SLPFs made a high contribution to the dietary patterns of subjects, showing a mean (SD) consumption of 5 ± 6%, while the consumption of SLPF meat, fish, and egg replacers made a very low contribution to their diet, with mean (SD) values of 1 ± 2% of total diet, which was a very low percentage of the total diet. Therefore, the results could be difficult to interpret. In this context, the dietary intake, anthropometric, and plasma biochemical parameters were only analysed considering the consumption of dairy SLPFs. In p = 0.02 and r = 0.31, p = 0.02, respectively). In addition, the ingested sugars through dairy SLPFs were also correlated with the plasma total cholesterol and LDL-C of subjects . Furthermore, the intake of total fats and carbohydrates from dairy SLPFs was correlated with plasma total cholesterol .Significant results were also observed in the plasma biochemical parameters. Subjects in the third tertile of dairy SLPFs had higher plasma total cholesterol and LDL-C than subjects in the two lowest tertiles, and these relationships were supported by positive correlations . It is worth highlighting that the two individuals who had high plasma levels of LDL-C also had higher consumption of dairy SLPFs.It is interesting to note that when these results were stratified by plasma reference values , particiAs far as we know, for the first time, this study compares the nutritional composition of Spanish SLPFs with that of normal protein foods. It is worth noting that, currently, there are no available databases in Spain of these specific products. In this regard, few studies have reported nutritional differences between SLPFs and normal protein foods ,11,12. IIn our study, we identified significant nutritional differences between SLPFs and regular foods, particularly in the categories of dairy products, meat, fish, and egg replacers. However, the findings of the previous study conducted by MacDonald et al. , which eTo our knowledge, this is the first study investigating the eating behaviour of children with IEMs and the impact of SLPFs on dietary patterns and biochemical profile, taking into account protein restrictions and how they could affect acquired and modifiable risk factors that exacerbate these hereditary conditions.SLPFs are an essential component of dietary treatment, contributing a significant portion of energy intake  in our patients. This is consistent with the findings of Daly et al. , who repp < 0.05), as well as a positive correlation between sugars in the total diet and sugars from dairy SLPFs . High sugar consumption has been linked to an increased likelihood of overweight and obesity among youth, regardless of other dietary or macronutrient intakes [p = 0.05; data not shown). However, despite calculating the contribution of SLPFs in the dietary pattern, Moretti et al. did not investigate the association of their consumption with plasma glucose metabolism or lipid profile parameters [The results of our study indicated that the percentage of total sugars consumed by the population exceeded the recommendations , althoug intakes  and alte intakes . In a st intakes . Interesrameters . On the rameters . Interesrameters  showed trameters . Howeverp = 0.03), highlighting another potential benefit of consuming these foods in reducing cardiovascular risk [F. prausnitzii, which is considered a biomarker of health status and a butyrate producer [On the other hand, our results suggest that fibre intake is adequate. This is mainly due to the fact that a significant proportion of our patients (68%) consume between 400 g and 600 g of fruits and vegetables per day, as recommended . The conlar risk . Despitelar risk . Bassaniproducer . In our producer . Foods cproducer . It is iproducer . In lighFurthermore, we observed that individuals who consumed more than 5% of their energy from dairy SLPFs also had higher dietary SFAs and sugars. These could be possible factors that potentially contribute to elevated levels of total cholesterol (TC), as well as LDL-C, in the blood. Thus, excessive consumption of SFAs, added sugars (such as sucrose or high fructose corn syrup), and refined carbohydrates leads to changes in LDL-C, HDL-C, and triglyceride, which can increase, and has been associated with increased morbidity and mortality from cardiovascular diseases, both in adults and children ,50,51,52However, our study has some limitations. One these is the incomplete nutritional information available for some SLPFs, including information about the kind of starch or fibre they contain. The other limitation is that our sample was not homogeneous for the different AA diseases, so it is not possible to compare the dietary pattern between them. The sample size is also limited; however, the statistical power for the main results is adequate since the estimated effect size for plasma total cholesterol levels assessed through dairy SLPF consumption indicated a statistical power of 86%. In addition, the estimated effect size for LDL-C assessed through dairy SLPF consumption revealed a statistical power of 83%. Moreover, the difficulty of working with the paediatric population and the low prevalence of these diseases make our findings particularly relevant and representative. Therefore, our study serves as an important example of the quality of the diet required for managing IEMs and highlights the need for further research in this area.Our results suggests that the nutritional composition of SLPFs is different from that of usual foods with protein and their contribution between 18% and 30% to the total diet of our participants has not been related to protein restriction, nor to the excessive consumption of SFAs or carbohydrates. However, a consumption of more than 5% of some SLPFs, such as dairy products, could imply the intake of more sugars, total fats, and SFAs, which could affect the cholesterol levels of these children. Our study highlights the importance of regularly evaluating the protein-controlled diet in IEM in clinical practice to ensure adequate intake of SLPFs and prevent imbalances that could impact the lipid and glucose profile of these patients. As a result, they could prevent potential future complications, including cardiovascular risk factors.