Search Results (17)

Alphaviruses engage a diverse array of attachment factors and receptors during viral entry, resulting in a broad host range and disease spectrum, and thus presenting them as a major global public health concern. The development of effective antivirals against these arboviruses relies on a comprehensive understanding of the molecular interplay between these viruses and host cell factors, as well as the wide range of immune responses that ensue following viral infection. In this review, we present the current understanding of the complex landscape of alphavirus interaction with attachment factors and entry receptors, some of which are characterized structurally, while others are characterized biochemically. Additionally, we provide an overview of the molecular bases of epitope recognition by neutralizing and non-neutralizing antibodies against alphaviruses, and how icosahedral symmetry influences these interactions, such as occupancy and neutralization potency. We further discuss the structural bases of epitope recognition of a few pan-alphavirus antibodies, their potential therapeutic implications, and offer future perspectives on the development of effective therapeutics against clinically relevant alphaviruses. Full article

Artificial intelligence (AI)-powered mental health chatbots have evolved quickly as scalable means for psychological support, bringing novel solutions through natural language processing (NLP), mobile accessibility, and generative AI. This systematic literature review (SLR), following PRISMA 2020 guidelines, collates evidence from 25 published, peer-reviewed studies between 2020 and 2025 and reviews therapeutic techniques, cultural adaptation, technical design, system assessment, and ethics. Studies were extracted from seven academic databases, screened against specific inclusion criteria, and thematically analyzed. Cognitive behavioral therapy (CBT) was the most common therapeutic model, featured in 15 systems, frequently being used jointly with journaling, mindfulness, and behavioral activation, followed by emotion-based approaches, which were featured in seven systems. Innovative techniques like GPT-based emotional processing, multimodal interaction (e.g., AR/VR), and LSTM-SVM classification models (greater than 94% accuracy) showed increased conversation flexibility but missed long-term clinical validation. Cultural adaptability was varied, and effective localization was seen in systems like XiaoE, okBot, and Luda Lee, while Western-oriented systems had restricted contextual adaptability. Accessibility and inclusivity are still major challenges, especially within low-resource settings, since digital literacy, support for multiple languages, and infrastructure deficits are still challenges. Ethical aspects—data privacy, explainability, and crisis plans—were under-evidenced for most deployments. This review is different from previous ones since it focuses on cultural adaptability, ethics, and hybrid public health incorporation and proposes a comprehensive approach for deploying AI mental health chatbots safely, effectively, and inclusively. Central to this review, symmetry is emphasized as a fundamental idea incorporated into frameworks for cultural adaptation, decision-making processes, and therapeutic structures. In particular, symmetry ensures equal cultural responsiveness, balanced user–chatbot interactions, and ethically aligned AI systems, all of which enhance the efficacy and dependability of mental health services. Recognizing these benefits, the review further underscores the necessity for more rigorous academic research into the development, deployment, and evaluation of mental health chatbots and apps, particularly to address cultural sensitivity, ethical accountability, and long-term clinical outcomes. Full article

Background/Objectives: Transcranial Direct Current Stimulation (tDCS) has proven to be a promising intervention for major depressive disorder (MDD). Even so, the specific neurophysiological mechanisms underlying its therapeutic effects, particularly regarding frontal EEG markers, remain insufficiently understood. This pilot study investigated both the clinical efficacy and neurophysiological impact of frontal tDCS in individuals with mild to severe depression, with particular focus on mood changes and alterations in Frontal Alpha Asymmetry (FAA), Beta Symmetry, and Theta/Alpha Ratios at the F3 and F4 electrode sites. Methods: A total of thirty–one participants were enrolled and completed a standardized Flow Neuroscience tDCS protocol targeting the dorsolateral prefrontal cortex using a bilateral F3/F4 montage. The intervention included an active phase of five stimulations per week for three weeks, followed by a Strengthening Phase with two stimulations per week. Clinical outcomes were assessed using the Montgomery–Åsberg Depression Rating Scale (MADRS), while neurophysiological changes were evaluated via standardized quantitative EEG (QEEG) recordings obtained before and after the treatment course. Among the participants, fourteen individuals had a baseline MADRS score of ≥20, indicating moderate to severe depressive symptoms. Results: Following tDCS treatment, significant reductions in MADRS scores were observed across the cohort, with clinical response rates notably higher in the moderate/severe group (71.4%) compared to the mild depression group (20.0%). Neurophysiological effects were modest: no significant changes were detected in FAA or Beta Symmetry measures. However, a substantial reduction in the Theta/Alpha Ratio at F4 was found in participants with moderate to severe depression (p = 0.018, Cohen’s d = −0.72), suggesting enhanced frontal cortical activation associated with clinical improvement. Conclusions: These findings indicate that frontal tDCS is effective in reducing depressive symptoms, particularly in cases of moderate to severe depression. While improvements in FAA and Beta Symmetry were not significant, changes in the Theta/Alpha Ratio at F4 point toward dynamic neurophysiological reorganization potentially linked to therapeutic outcomes. The Theta/Alpha Ratio may serve as a promising biomarker for tracking tDCS response, whereas other EEG metrics might represent more stable trait characteristics. Future research should prioritize individualized stimulation protocols and incorporate more sensitive neurophysiological assessments, including functional connectivity analyses and task-evoked EEG paradigms, to understand the mechanisms underlying clinical improvements. Full article

Background: Defined as a self-selected speed of <0.4 m/s, chronic stroke survivors falling in this category are classified as “severe”, usually homebound and sedentary, and they experience worse outcomes. Limited rehabilitation strategies are available to improve walking speed and related outcomes in this subgroup, and questions regarding effective rehabilitation options remain. The objective of this study was to determine the effects of backward (BLTT) and forward (FLTT) locomotor treadmill training on overground walking speed, spatiotemporal symmetry, and dynamic postural stability. Methods: In this single-center, assessor-blinded, randomized controlled pilot trial, 14 stroke survivors with severe waking impairment underwent 12 sessions of BLTT (n = 7) or FLTT (n = 7). The primary outcome was the proportion of participants reaching clinically meaningful important difference (MCID) on the 10-meter walk test following training completion. Secondary outcomes were between-group differences in walking speed, spatiotemporal symmetry, and completion time on the 3-meter timed up and go (3M TUG) at 24 h, 30 days, and 90 days POST. Results: Two subjects in the BLTT group (28.6%) and one (14.3%) in FLTT achieved MCID following training; however, most subjects did not, with significant variability in response. At 24 h POST, the median (IQR) percent change in walking speed was 28.9 (9.01–36.7) and 17.4 (12.6–39.7) with BLTT and FLTT, respectively; however, no between-group differences were seen (p = 0.80) at this time point or at 30 (p > 0.99) and 90 (p > 0.99) days follow up. Likewise, there were no significant between-group differences in spatiotemporal symmetry and the 3M TUG across time points. Conclusions: While preliminary, this study found that 12 training sessions did not lead to group-level achievement of MCID for walking speed in our cohort and found no significant between-group differences in walking capacity or dynamic postural stability. Future well-powered dosing trials and mechanistically driven studies are needed to optimize and identify predictors of training response. Full article

Severe traumatic brain injury (sTBI) often results in significant impairments in gait stability, symmetry, and smoothness. Inertial measurement units (IMUs) have emerged as powerful tools to quantify these aspects of gait, but their clinometric properties in sTBI populations remain underexplored. This study aimed to assess the test-retest reliability and minimal detectable change (MDC) of three IMU-derived indices—normalized Root Mean Square (nRMS), improved Harmonic Ratio (iHR), and Log Dimensionless Jerk (LDLJ)—during a 10 m walking test for sTBI survivors. Forty-nine participants with sTBI completed the walking test, with IMUs placed on key body segments to capture accelerations and angular velocities. Test-retest analyses revealed moderate to excellent reliability for nRMS and iHR in anteroposterior (ICC: 0.78–0.95 and 0.94, respectively) and craniocaudal directions (ICC: 0.95), with small MDC values, supporting their clinical applicability (MDC: 0.04–0.3). However, iHR in the mediolateral direction exhibited greater variability (ICC: 0.80; MDC: 9.74), highlighting potential sensitivity challenges. LDLJ metrics showed moderate reliability (ICC: 0.57–0.77) and higher MDC values (0.55–0.75), suggesting the need for further validation. These findings underscore the reliability and sensitivity of specific IMU-derived indices in detecting meaningful gait changes in sTBI survivors, paving the way for refined assessments and monitoring the rehabilitation process of sTBI survivors. Future research should explore these indices’ responsiveness to interventions and their correlation with functional outcomes. Full article

Background/Objectives: Atypical parkinsonian syndromes (APSs) are a group of neurodegenerative disorders that differ from idiopathic Parkinson’s disease (IPD) in their clinical presentation, underlying pathology, and response to treatment. APSs include conditions such as multiple system atrophy (MSA), progressive supranuclear palsy (PSP), corticobasal syndrome (CBS), and dementia with Lewy bodies (DLB). These disorders are characterized by a combination of parkinsonian features and additional symptoms, such as autonomic dysfunction, supranuclear gaze palsy, and asymmetric motor symptoms. Many hypotheses attempt to explain the causes of neurodegeneration in APSs, including interactions between environmental toxins, tau or α-synuclein pathology, oxidative stress, microglial activation, and vascular factors. While extensive research has been conducted on APSs, there is a limited understanding of the symmetry in these diseases, particularly in MSA. Neuroimaging studies have revealed metabolic, structural, and functional abnormalities that contribute to the asymmetry in APSs. The asymmetry in CBS is possibly caused by a variable reduction in striatal D2 receptor binding, as demonstrated in single-photon emission computed tomography (SPECT) examinations, which may explain the disease’s asymmetric manifestation and poor response to dopaminergic therapy. In PSP, clinical dysfunction correlates with white matter tract degeneration in the superior cerebellar peduncles and corpus callosum. MSA often involves atrophy in the pons, putamen, and cerebellum, with clinical symmetry potentially depending on the symmetry of the atrophy. The aim of this review is to present the study findings on potential symmetry as a tool for determining potential neuropsychological disturbances and properly diagnosing APSs to lessen the misdiagnosis rate. Methods: A comprehensive review of the academic literature was conducted using the medical literature available in PubMed. Appropriate studies were evaluated and examined based on patient characteristics and clinical and imaging examination outcomes in the context of potential asymmetry. Results: Among over 1000 patients whose data were collected, PSP-RS was symmetrical in approximately 84% ± 3% of cases, with S-CBD showing similar results. PSP-P was symmetrical in about 53–55% of cases, while PSP-CBS was symmetrical in fewer than half of the cases. MSA-C was symmetrical in around 40% of cases. It appears that MSA-P exhibits symmetry in about 15–35% of cases. CBS, according to the criteria, is a disease with an asymmetrical clinical presentation in 90–99% of cases. Similar results were obtained via imaging methods, but transcranial sonography produced different results. Conclusions: Determining neurodegeneration symmetry may help identify functional deficits and improve diagnostic accuracy. Patients with significant asymmetry in neurodegeneration may exhibit different neuropsychological symptoms based on their individual brain lateralization, impacting their cognitive functioning and quality of life. Full article

Background: The side hop test (SHT) measures the number of jumps performed over 30 s. Although this measure has demonstrated its value in clinical practice, the temporal parameters of the SHT allow for a deeper analysis of the execution strategy. The aim of this study is to assess the reliability and construct validity of contact time parameters during the SHT recorded by a video analysis system in anterior cruciate ligament reconstructed (ACLR) patients. Methods: We investigated the reliability (intra-rater, standard error of measurement (SEM), and minimum detectable change (MDC)), discriminant validity (operated (OP) versus non-operated (NOP) side), and convergent validity (relationship with strength and psychological readiness) of SHT contact time parameters, number of valid hops and limb symmetry index (LSI) in 38 ACLR patients. Contact time parameters are presented as mean, standard deviation (SD), and coefficient of variation (CV) of contact time. Results: Intra-tester reliability was good to excellent for all contact time parameters. For discriminant validity, the mean and SD contact times of the OP leg were significantly longer than those of the NOP leg, although the difference was smaller than the SEM and MDC values. The number of valid jumps and CV contact time parameters were not significantly different. Isokinetic quadriceps strength (60°/s) was strongly correlated with mean contact time for both legs. However, psychological readiness was not correlated with any of the contact time parameters. Conclusions: Temporal parameters of the SHT measured on video analysis are valid and reliable parameters to assess the performance strategy of the SHT. The results should be interpreted with caution regarding the SEM and MDC values. Further studies are needed to measure criterion validity, inter-rater reliability, and responsiveness. Full article

In this retrospective study to determine the optimal method of evaluating static weight-bearing distribution to assess response to treatment in dogs with osteoarthritis using a weight distribution platform, data from the Clinica Veterinária de Cães (Portuguese Republican National Guard) clinical records were extracted. At baseline and at 15 days post-treatment, follow-up data from 80 dogs treated for bilateral hip osteoarthritis were categorized based on an anchor question. Estimates of minimal clinically important differences were calculated with distribution-based and anchor-based methods for deviation from normal weight-bearing and a symmetry index (SI). For deviation, the anchor-based methods provided a range from −0.3 to −3.1, and the distribution-based methods from 0.16 to 0.29. For SI, the anchor-based methods provided a range from −10.0 to −23.9, and the distribution-based methods from 1.31 to 2.88. Receiver operator characteristic curves provided areas under the curve >0.7, indicating an acceptable cut-off point. We presented estimates of −1 for deviation and −10 for SI in dogs with OA. These estimates can be used for research and patient monitoring. Future studies should include OA from other joints and animals from a broader clinical context. Full article

This umbrella review aimed to determine the effectiveness of nonpharmacological interventions in pulmonary ventilation and their impact on respiratory function. An individual with impaired ventilation displays visible variations manifested in their respiratory frequency, breathing rhythm ratio (I:E), thoracic symmetry, use of accessory muscles, dyspnea (feeling short of breath), oxygen saturation, diaphragm mobility, minute ventilation, peak flow, walking test, spirometry, Pimax/Pemax, diffusion, and respiratory muscle strength. Any variation in these markers demands the need for interventions in order to duly manage the signs and symptoms and to improve ventilation. Method: Systematic reviews of the literature published in English, Spanish, French, and Portuguese were used, which included studies in which nonpharmacological interventions were used as a response to impaired ventilation in adults in any given context of the clinical practice. The recommendations given by the Joanna Briggs Institute (JBI) for umbrella reviews were followed. This research took place in several databases such as MEDLINE, CINAHL Complete, CINHAL, MedicLatina, ERIC, Cochrane Reviews (Embase), and PubMed. The Joanna Briggs critical analysis verification list was used for the systematic review. The data extraction was performed independently by two investigators based on the data extraction tools of the Joanna Briggs Institute, and the data were presented in a summary table alongside the support text. Results: Forty-four systematic reviews, thirty randomized clinical essays, and fourteen observational studies were included in this review. The number of participants varied between n = 103 and n = 13,370. Fifteen systematic revisions evaluated the effect of isolated respiratory muscular training; six systematic revisions evaluated, in isolation, breathing control (relaxed breathing, pursed-lip breathing, and diaphragmatic breathing exercises) and thoracic expansion exercises; and one systematic review evaluated, in isolation, the positions that optimize ventilation. Nineteen systematic reviews with combined interventions that reinforced the role of education and capacitation while also aiming for their success were considered. The articles analyzed isolated interventions and presented their efficacy. The interventions based on respiratory exercises and respiratory muscular training were the most common, and one article mentioned the efficacy of positioning in the compromisation of ventilation. Combined interventions in which the educational component was included were found to be effective in improving pulmonary function, diffusion, oxygenation, and functional capacity. The outcomes used in each study were variable, leading to a more difficult analysis of the data. Conclusions: The interventions that were the focus of the review were duly mapped. The results suggest that nonpharmacological interventions used to optimize ventilation are effective, with a moderate to high level of evidence. There is a strong foundation for the use of the chosen interventions. The lack of studies on the intervention of “positioning to optimize ventilation” points out the need for a deeper analysis of its effects and for studies with a clear focus. This study supports the decisions and recommendations for the prescription of these interventions to patients with impaired ventilation. Full article

Osteoarthritis (OA) is the most commonly diagnosed joint disease in companion animals, and proper tools are necessary to assess patients and response to treatment. We aimed to perform the psychometric evaluation of several clinical metrology instruments (CMI), developed to evaluate pain and assess outcome. Fifty police working dogs with bilateral hip OA were assessed in a prospective, randomised, double-blinded study. Patients were evaluated using a stance analyser in six different moments divided over a 180-day period. Pedometer step count, weight-bearing symmetry index and deviation from normal weight-bearing were calculated and used for criterion validity. In each evaluation moment, a copy of the Hudson Visual Analogue Scale (HVAS), Canine Brief Pain Inventory (CBPI), Liverpool Osteoarthritis in Dogs (LOAD) and Canine Orthopaedic Index (COI) were completed by the dogs’ handlers. Correlations between CMIs were evaluated as construct validity. Further evaluation was performed with the Kaiser–Meyer–Olin measure of sampling adequacy, Eigenvalue and scree-plot analysis. Internal consistency was tested with Cronbach’s α. Significant weak correlation was found between all CMIs and stance analysis symmetry index measure and deviation, indicating criterion validity. Significant weak correlation was also found between pedometer count and LOAD plus COI. Cronbach’s α was 0.80 for HVAS, 0.98 for CBPI, 0.97 for LOAD and 0.98 for COI. Significant strong correlation was observed between CMIs, indicating construct validity. We present criterion and construct validity of these CMIs, which are able to capture various dimensions of OA. They can be used for the evaluation of osteoarthritis and response to treatment in dogs. Full article

In cerebral palsy (CP) the basis for rehabilitation comes from neuroplasticity. One of the leading therapeutic approaches used in the management of CP is the NDT Bobath therapy and Vojta therapy consists in trying to program the ideal movement patterns for the age. The aim of our research was to analyze, from a functional point of view, the evolution of the biomechanical parameters characterizing the balance, in children with CP. The group of 12 subjects average age of 7 ± 3.28 years. The subject’s evaluation included a functional clinical evaluation by Berg pediatric scale and a biomechanical evaluation performed using the “Stabilometry footboard PoData 2.00” for evaluation the body weight distribution on the foot level. The rehabilitation program was developed based on two methods, NDT Bobath and Vojta. A 90-min physiotherapy session starts with a Vojta therapy activation, for 20 min. Between the two therapies there is a 10-min break, then the session continues with NDT Bobath exercises within the 3 physical exercises proposed for 60 min. 5 days per week, 6 months. The analysis of the data collected before and after the application of the rehabilitation program, regarding the using the Berg scale indicates a progress of 32.35%, (p = 0.0001 < 0.05) and the effect size is large. The evolution of the data that indicate the distribution of body weight at the level of the two lower limbs, at the two moments pre/post, evaluation. For left side a progress of 8.39%, (p = 0.027 < 0.05) but a small effect size of 0.86. For right side a progress of 10.36% (p = 0.027 < 0.05) and also a small effect size of 0.86. Analyzing the results, we find that there is a left-right rebalancing in most patients. The favorable results that were obtained by drawing up a physiotherapy program composed of the combination of the two Vojta and NDT Bobath methods are proof of the fact that both methods are based on the creation of a stimulating peripheral pressure, which, if maintained, generates an extended stereotyped motor response. A pattern of symmetrical muscle contraction is thus created and thus balance and postural control can be achieved. The left-right rebalancing, proven by the percentage distribution analysis of the weight at the lower segmental level, demonstrated that the body alignment approach through the Vojta method on the one hand and the inhibitory facilitating postures/exercises promoted by the NDT Bobath method, allows obtaining a symmetry. Full article

Total knee replacement (TKR) is the treatment of choice for advanced stages of osteoarthritis but it requires good postoperative rehabilitation. This study aimed to assess the effectiveness of exercises using virtual reality to improve gait parameters in patients after TKR. Fifty-nine patients 7–14 days after TKR surgery were divided into a study group (VRG, n = 38) and a control group (CG, n = 21). Both groups underwent the same 4-week rehabilitation protocol. The VRG group had 12 additional nonimmersive virtual reality game sessions on the Virtual Balance Clinic prototype system at 30 min each, focusing on gait and balance improvement. Spatiotemporal, force and foot plantar pressure parameters were collected on an instrumented treadmill during a 30 s walk. The most significant improvement was in the symmetry indices of forefoot force, maximum forefoot force, loading response time, and preswing time (p < 0.05) in both groups. Gait speed increased by 31.25% and 44% in the VRG and CG groups, respectively (p < 0.005). However, the extra exergaming sessions did not significantly improve rehabilitation outcomes. Therefore, additional VR training does not improve gait better than standard rehabilitation alone, but the improvement of gait, especially its symmetry, is significant within the first six weeks after surgery. Full article

Walking function disorders are typical for patients after cerebral stroke. Biofeedback technology (BFB) is currently considered effective and promising for training walking function, including in patients after cerebral stroke. Most studies recognize that BFB training is a promising tool for improving walking function; however, the data on the use of highly selective walking parameters for BFB training are very limited. The aim of our study was to investigate the feasibility of using BFB training targeting one of the basic parameters of gait symmetry—stance phase duration—in cerebral stroke patients in the early recovery period. The study included 20 hemiparetic patients in the early recovery period after the first hemispheric ischemic stroke. The control group included 20 healthy subjects. The BFB training and biomechanical analysis of walking (before and after all BFB sessions) were done using an inertial system. The mean number of BFB sessions was nine (from 8 to 11) during the three weeks in clinic. There was not a single negative response to BFB training among the study patients, either during the sessions or later. The spatiotemporal parameters of walking showed the whole syndrome complex of slow walking and typical asymmetry of temporal walking parameters, and did not change significantly as a result of the study therapy. The changes were more significant for the functioning of hip and knee joints. The contralateral hip amplitude returned to the normal range. For the knee joint, the amplitude of the first flexion increased and the value of the amplitude of hyperextension decreased in the middle of the stance phase. Concerning muscle function, the observed significant decrease in the function of m. Gastrocnemius and the hamstring muscles on the paretic side remained without change at the end of the treatment course. We obtained positive dynamics of the biomechanical parameters of walking in patients after the BFB training course. The feasibility and efficacy of their use for targeted correction need further research. Full article

This study aims to analyze and compare the disease activity control and quality of life of patients with rheumatoid arthritis (RA) who were treated with biological products in real-life settings. We tried to determine whether there is a symmetry in the performance of the biological molecules between each other and with the first Janus kinase (JAK) inhibitor. This is an observational, longitudinal, real-life study performed in the biggest rheumatology clinic during the period 2012–2020 comparing quality of life, cost of therapy, and disease control via different clinical measures. In all three disease activities measurement instruments, we observed an improvement for all biologic and target synthetic medicines. The disease activity score (DAS28) score decreased from 5.06 to 3.01, on average, for all INNs, suggesting that the majority of patients move away from moderate to low disease activity. The clinical disease activity index (CDAI) score decreased from 25.9 to 9.4, also indicating that patients with moderate disease activity reached a low level of activity. Similar results are reflected in the score, which fell from 27.7 to 10.3, again confirming the improvement to a low level of disease activity for patients treated with all INNs. Logically, with the successful control of disease activity, the quality of life (QoL) of the observed patients improved from 0.77 to 0.83 after a one-year follow up, as measured with the EuroQuol 5D-3L (EQ5D). Based on these results, we can consider that the observed biological INNs perform symmetrically in terms of the control of disease activity and improvement in the QoL of the observed patients. Biological therapy improves the disease control and quality of life of suitable patients with RA in real-life settings. All available biological therapies could be used interchangeably. Full article

Parvovirus B19 (B19V), an ssDNA virus in the family Parvoviridae, is a human pathogenic virus, responsible for a wide range of clinical manifestations, still in need of effective and specific antivirals. DNA structures, including G-quadruplex (G4), have been recognised as relevant functional features in viral genomes, and small-molecule ligands binding to these structures are promising antiviral compounds. Bioinformatic tools predict the presence of potential G4 forming sequences (PQSs) in the genome of B19V, raising interest as targets for antiviral strategies. Predictions locate PQSs in the genomic terminal regions, in proximity to replicative origins. The actual propensity of these PQSs to form G4 structures was investigated by circular dichroism spectroscopic analysis on synthetic oligonucleotides of corresponding sequences. No signature of G4 structures was detected, and the interaction with the G4 ligand BRACO-19 (N,N′-(9-{[4-(dimethylamino)phenyl]amino}acridine-3,6-diyl)bis(3-pyrrolidin-1-ylpropanamide) did not appear consistent with the stabilisation of G4 structures. Any potential role of PQSs in the viral lifecycle was then assessed in an in vitro infection model system, by evaluating any variation in replication or expression of B19V in the presence of the G4 ligands BRACO-19 and pyridostatin. Neither showed a significant inhibitory activity on B19V replication or expression. Experimental challenge did not support bioinformatic predictions. The terminal regions of B19V are characterised by relevant sequence and symmetry constraints, which are functional to viral replication. Our experiments suggest that these impose a stringent requirement prevailing over the propensity of forming actual G4 structures. Full article