Rapid correction of hyponatremia is the most frequent predisposing factor for the development of central pontine myelinolysis (CPM). Alcoholism, cirrhosis, malnutrition, and severe burns are associated conditions that often present in combination with a rapid rise in serum sodium concentration. However, its association with hyperglycemia has not been as well established. There have been recent reports of acute to subacute presentation of CPM with hyperglycemia. We report an unusual case of a 48-year-old Caucasian male who presented with pseudobulbar palsy, ataxia, and quadriplegia with worsening pontine hyperintensities and was diagnosed with CPM in the setting of persistent hyperglycemia with normal serum sodium. Central pontine myelinolysis (CPM), a condition described in 1959 by Adams et al., was first associated with young alcoholics . HyperosA 48-year-old Caucasian male presented with generalized limb weakness and incoordination of about one-month duration. He also complained of excessive thirst and eating. At the time of presentation, no focal neurologic deficit was seen on examination. Blood glucose was 608 mg/dL with a corrected sodium level of 133 mmol/L and normal serum potassium of 3.8 mmol/L. His hyperglycemia was treated. Glycated hemoglobin was 14.7%. He had no prior history of diabetes mellitus before this presentation. He had magnetic resonance imaging (MRI) of the brain, which showed signal intensity in central pons on T2-weighted images , normal serum sodium of 136 mmol/L, normal potassium of 3.7 mmol/L, and glycated hemoglobin of 12.4%. The initial working diagnosis was a motor neuron disease with the upper and lower motor neuron signs; however, electromyography was done, which showed no evidence of motor neuron disease, and a mild predominant sensory axonal length-dependent polyneuropathy was seen, which is likely due to his uncontrolled diabetes mellitus. The fasciculations were thought to be due to diabetic neuropathy.MRI of the brain showed worsening signal intensity in the central pons on T2-weighted and fluid-attenuated inversion recovery (FLAIR) sequences Figures . There wThe term “central pontine myelinolysis” was first described by Adams in 1959 as a symmetric, demyelinating focus prominent in the central pons . ChronicIn 1988, McKee et al. concluded based on postmortem diagnoses that extreme serum hyperosmolality contributed to CPM, which was due to hypernatremia, hyperglycemia, and azotemia, alone or combined . HyponatThe pons, which is primarily affected, was proposed to be due to an extensive gray-white matter admixture providing a suitable environment for interaction between the richly vascularized gray matter and myelin-containing white matter . In 1982Despite all of the cases where CPM has been reported, there are fewer reports of subacute and chronic presentations of hyperglycemia . In thisClinical features that have been described include quadriparesis, bulbar disorders, pathological reflexes, pseudobulbar emotionality, incontinence, decreased alertness, and confusion . In thisImaging findings of CPM include symmetric signal intensity abnormality in the central pons at T2-weighted and FLAIR imaging , which mWhile there have been several associations with CPM, such as alcoholism and rapid correction of hyponatremia, hyperglycemia is one of the less recognized associations. Recent evidence points to an association with hyperglycemia beyond pseudohyponatremia as the hyperosmolar state can also induce demyelination in the pons. Therefore, a diagnosis of CPM should be considered in diabetic patients with poor glycemic control.


Robotic developments in the field of rehabilitation and assistance have seen a significant increase in the last few years. The improvement of biosensing technologies provides robust ways of assessing the user’s neuromechanical behavior and mental states, which, when combined with robotic therapies, leads to critical improvements in motor or cognitive function recovery. Recent advances in bioelectrical signal processing and acquisition devices, in computer-vision techniques and machine-learning, and in the kinetic and dynamic analysis of movement have had a huge impact on the efficient development of the aforementioned robotic approaches.This Special Issue is focused on breakthrough developments in the field of biosensors applied to rehabilitation and assistive robotics. The Special Issue has collected five outstanding papers covering different aspects of biosensing technologies and their use in different environments. What follows is a summary of the scope and main contributions of each of these papers, provided as a teaser for the interested reader.The use of biosensors to monitor and assess progress during rehabilitation therapies is a key tool in many recent studies. For instance, the performance of a robotic ankle orthosis T-FLEX in combination with a lower-limb exoskeleton AGoRA has been evaluated using electromyographic (EMG) sensors to study muscular activity and using inertial sensors to study gait patterns . Other pAnother important application of biosensors in assistive technologies is their use as a way of inferring patients’ actual or intended motion to control external devices. A way of achieving this is by processing cortical activity with non-invasive electroencephalographic (EEG) devices. In , this ap


A two-way repeated-measures analysis of variance was used to test the null hypothesis of no different change in VAS over time between groups. After the intervention, the MT group perceived largely greater psychological readiness (p < 0.01). MT and CON groups experienced a large reduction in VAS after the intervention (p < 0.001). However, a small time × group interaction was observed (p = 0.023). MT reported a greater perception of the psychological readiness of the soccer players and a lower perception of pain than those who performed conventional therapy.Virtual reality-guided imagery (VRGI) and mirror therapy (MT) have been used in isolation to treat patients suffering from different injuries. However, no attempts have been made to understand the effects of combined VRGI and MT added to conventional physical therapy, and no information exists regarding perceptual responses to these rehabilitation strategies in female football players. Thus, this study aimed to examine the effect of MT added to conventional rehabilitation on psychological readiness and perception of pain in ACL-injured female football players. Thirty ACL-injured female football players competing in the 2nd and 3rd Italian tier who underwent an ACL rehabilitation program from the same clinic participated in this study. Players were randomly distributed in an MT group (n = 15) and a CON group (n = 15). All participants reported their perception of pain on a VAS before and after the interventions and their psychological readiness to return to sport after ACL injury and reconstruction surgery on the ACL-RSI scale after the intervention. An independent-sample  The knee, with special emphasis on the anterior cruciate ligament (ACL), is often cited as the most common injury site in female football players ,2,3,4. FPsychological responses to specific rehabilitation programs have been identified as a potentially modifiable factor associated with knee function ,12,13. IInjured athletes can benefit from mental practice, such as hypnosis, meditation, goal setting, self-talk, and imagery . For exaIn summary, both VRMI and MT have been used in isolation to treat patients suffering from different injuries. However, no attempt has been made to understand the effects of combined VRMI and MT added to conventional physical therapy, and no information exists regarding perceptual responses to these rehabilitation strategies in female football players. Thus, this study aimed to examine the effect of MT added to conventional rehabilitation on psychological readiness and perception of pain in ACL-injured female football players.A total of 30 ACL-injured female football players competing in the 2nd and 3rd Italian tier who underwent an ACL rehabilitation program from the same clinic participated in this study. Inclusion criteria were an age of >18 years old and a minimum of 3 years competing on the first team at the national level. All procedures followed the guidelines of the Declaration of Helsinki and were in line with established ethical standards in sports sciences . WrittenThe participants were recruited throughout a 5-year period, as each player underwent ACL rehabilitation at different times. Players were randomly distributed into an MT group  and a control group . Both MT and CON interventions lasted 20 weeks. All participants reported pain perception in a VAS before and after the interventions and their psychological readiness in an ACL-RSI after the intervention.Early rehabilitation (1 to 10 weeks): the objectives in this phase are the recovery of the ROM of the injured joint and controlled walking;Advanced mobilization (from week 11 to 16): the objectives are the complete recovery of the ROM and the joint and muscle component;Reconditioning (weeks 17 to 20): the objectives are to improve walking and specific sports exercises for returning to the field.The CON group underwent a 20-week conventional physical therapy program. Training frequency was 5/week with a session length of 60 min. The rehabilitation program was divided into three stages, specifically: A certified physiotherapist, based on the acute responses of the patients, adjusted the intensity of the exercise. The conventional physical therapy used in this study comes from the Brotzman and Wilk’s method .Relaxation phase. Typically consists of 3–5 min of relaxation, in which the person is asked to imagine himself in a warm and relaxing place  and to contract and relax the muscles. Adding a relaxation period should improve concentration, promote a more vivid motor image, and improve attention and performance.Nuclear phase. 10–15 min follow-up in which suggestions are given for imagining internal images related to the use of the affected limb in one or more functional tasks and also through the mirror box.Final Phase. The recording ends with 3 to 5 min of concentrating on what has been done previously.The MT group underwent a 20-week conventional physical therapy program plus MT. Training frequency was 5/week with a session length of 60 min, with MT added 3 times/week for 20–25 min after the end of the conventional physical therapy. The MT consisted of the following:The teaching tool used is the exercise of mental representation.Work with exercises in the healthy limb mirror box .-Mirror box . The following presents a series of exercises that were performed during the 1–10-week period:Quadriceps isometric contraction exercises (patient sitting);Flexion/extension of the ankle (patient sitting and standing);Knee flexion/extension (patient sitting and standing);Hip flexion/extension/circling (sitting and standing);Limb adduction/abduction/circling (sitting and standing).-Motor (10 views/1 motor repetition x 5 series increasing to 10) from weeks 11–16:Front/side/back lunge (standing);Single-leg squat (standing);Bipodalic squat (standing);Ascent and descent of steps (standing).-Motor (10 views/1 motor repetition x 5 sets increasing to 10) from weeks 17–20:Stroke ;Stop ;Passage ;Jump to hit the head;Guide the ball;Parade.The sets and repetitions vary according to the patient, their health, and pain perception. Specifically, the mental practice includes:At the beginning (week 0) and the end of the treatment (week 20), participants were asked to complete the 0 to 10 Likert scale again (anchored to two endpoints labeled “no pain” and “worst pain someone could ever experience”) to report the pain experienced. This scale is a valid, internally consistent, and reliable measure of pain in different populations .At the end of the rehabilitation (week 20), the patients scored their perception of psychological readiness to return to sport after ACL injury and reconstruction surgery on the ACL-RSI scale. The scale was anchored on a 0 to 100 AU and consisted of 12 items, which included 5 related to emotions, 5 to confidence in performance, and 2 to risk appraisal. Emotion-related questions included (1) Are you nervous about playing your sport? (2) Do you find it frustrating to have to consider your knee concerning your sport? (3) Do you feel relaxed about playing your sport? (4) Do you fear reinjuring your knee by playing your sport? (5) Are you afraid of accidentally injuring your knee by playing your sport? Questions related to confidence in performance included (6) Are you confident that your knee will not give way by playing your sport? (7) Are you confident that you could play your sport without concern for your knee? (8) Are you confident about your knee holding up under pressure? (9) Are you confident that you can perform at your previous level of sport participation? (10) Are you confident about your ability to perform well at your sport? Questions related to risk appraisal were (11) Do you think you are likely to reinjure your knee by participating in your sport? and (12) Do thoughts of having to go through surgery and rehabilitation again prevent you from playing your sport? This scale was found to have high internal consistency , and a fair-to-good predictive ability for a 12-month return-to-sport outcome  .p > 0.05). An independent sample t-test was performed to assess between-group differences in post-intervention ACL-RSI. A further independent sample t-test was performed to assume non-significant differences between VAS values before the intervention. A two-way repeated-measures analysis of variance (ANOVA) was used to test for differences in training-induced changes in VAS. The independent variables included one between-subjects factor (training intervention) with two levels (MT and CON) and one within-subject factor (time) with two levels (pre- and post-intervention). To examine the influence of training intervention on the development of our dependent variable, we used this ANOVA to test the null hypothesis of no different change over time between groups (training intervention × time interaction). To qualitatively interpret the magnitude of differences, effect sizes (d), and associated, 95% confidence intervals (95% CI) were classified as small (0.2–0.5), moderate (0.5–0.8), and large (>0.8) [p ≤ 0.05. Data analyses were performed using Statistical Package for Social Science software .A Shapiro-Wilks test revealed that ACL-RSI and VAS values were normally distributed within each group at evaluation moments (e (>0.8) . Descripp < 0.01; p < 0.001; p = 0.023).A detailed description of participants’ ACL-RSI scores is reported in The main finding of this pilot study was that female football players undergoing MT-based rehabilitation reported a greater perception of psychological readiness compared to those involved in conventional physical therapy. Both groups experienced a similar reduction in pain perception after the rehabilitation. Undergoing ACL rehabilitation sooner after injury, along with a high motivation to return to sports, may impact a player’s return to play after an ACL injury .An injury weakens an athlete’s physical ability, but above all there is the psychological aspect. According to a study, 55% of athletes resume playing sports after surgery, but at a lower level than that practiced before getting hurt . TherefoUnderstanding the relationship between psychological factors and returning to sports is essential because most athletes recover good physical function after surgery, yet, many athletes with good knee function do not return to sports . For exaThis study has some limitations. First, the sample consists of a small number of participants. Second, the protocol administered did not follow a validation procedure. Finally, pain perception should have been measured at each therapy session; instead, in our case, it was measured only at the beginning and end of treatment. Given these limitations, there is a need to replicate the study to extend beyond the generalization of the results obtained.Mirror therapy reported a greater perception of the psychological readiness of the soccer players and a lower perception of pain compared to those who performed conventional therapy. Although both groups experienced a similar reduction in pain perception after rehabilitation, those who performed mirror therapy felt more ready to return to sports. It is important to consider the psychological aspect concerning players, as the fear of new injuries can affect their return to sport. For Mirror Therapy to become a standard module of treatment in sports rehabilitation, there is a need for collaboration between the physical therapist and other staff professionals, such as the mental coach, sports psychologist, osteopath, and orthopedic surgeon. They could define a protocol to improve function and perceived quality of life, not only in the rehabilitation treatment of athletes in the post-acute but also in the chronic phase. The total duration of the treatment could vary from 1 week to 8 weeks. Fifty percent of the treatments have an intervention period of 4 weeks , with a


N = 31) and people with bipolar disorder , first-degree biological relatives of these patient groups , and healthy controls (N = 29). PSZ exhibited reduced surround suppression across orientations; although group differences were minimal at the condition that produced the strongest suppression. PBD and PSZrel exhibited intermediate suppression, whereas PBDrel performed most similarly to controls. Intriguingly, group differences in orientation-dependent surround suppression magnitude were moderated by visual acuity. A simulation in which visual acuity and/or focal attention interact with untuned gain control reproduces the observed pattern of results, including the lack of group differences when orientation of center and surround are the same. Our findings further elucidate perceptual mechanisms of impaired center-surround processing in psychosis and provide insights into the effects of visual acuity on orientation-dependent suppression in PSZ.Perceptual distortions are core features of psychosis. Weakened contrast surround suppression has been proposed as a neural mechanism underlying atypical perceptual experiences. Although previous work has measured suppression by asking participants to report the perceived contrast of a low-contrast target surrounded by a high-contrast surround, it is possible to modulate perceived contrast solely by manipulating the orientation of a matched-contrast center and surround. Removing the bottom-up segmentation cue of contrast difference and isolating orientation-dependent suppression may clarify the neural processes responsible for atypical surround suppression in psychosis. We examined surround suppression across a spectrum of psychotic psychopathology including people with schizophrenia (PSZ;  Perceptual distortions are a primary symptom of psychosis. In 2005, Several neural mechanisms – occurring both inside (i.e. intrinsic) and outside (i.e. extrinsic) of primary visual cortex (V1) work together to determine perceived contrast, which can generally be predicted from firing rates of neurons in V1 . One sucFeedback from higher areas in the visual cortex is also known to alter firing rates of neurons in primary visual cortex . In partThe present study implemented a novel contrast-matched surround suppression paradigm that manipulated relative center-surround orientation  and distance . The goal was to collect the behavioral data necessary to determine whether perceptual contrast surround suppression deficits associated with schizophrenia might be attributed solely to orientation-insensitive, V1-intrinsic suppression mechanisms (untuned gain control) or whether orientation-dependent mechanisms are also altered. In the event that we observed alterations of orientation-dependent suppression, we included a far-surround condition to enable further discrimination between V1-intrinsic and V1-extrinsic mechanisms. The surround receptive field can be disaggregated into two regions (termed near and far surround) that are mediated by different neural mechanisms. The far surround is mediated by feedback connections extrinsic to V1, whereas the near-surround is mediated by a combination of both feedback connections and V1-intrinsic horizontal connections . Thus, bTypically, contrast surround suppression is measured with a low-contrast target embedded in a high-contrast surround, but, in the present study, the luminance contrasts of the center and surround gratings were matched. Although this choice reduces the expected magnitude of the behavioral effect , it alsoIt is unclear whether atypical surround suppression is present in other psychotic disorders, such as bipolar disorder . Given tBased on previous work with a similar transdiagnostic outpatient sample , we hypoPatients were recruited from Minneapolis Veterans Affairs Health Care System (MVAHCS) outpatient clinics, community support programs for the mentally ill, and county mental health clinics. First degree relatives of PSZ and PBD were identified by research staff using a pedigree form completed through interviews with patients and were invited by mail and telephone to participate in the study. Healthy controls (HCs) were recruited via posted announcements at fitness centers, community libraries, the MVAHCS, and newsletters for veterans. Potential PSZ, PBD, and HC were excluded if they met any of the following criteria: English as a second language, age >60 years, IQ <70, substance dependence within the past 6 months, substance abuse within 2 weeks of testing, head injury with skull fracture or substantial loss of consciousness (i.e. loss of consciousness >30 minutes), electroconvulsive therapy, amblyopia untreated before 18, epilepsy, stroke, or other neurological conditions. Additional exclusion criteria for HC were first-degree family history of major depressive disorder or a psychotic disorder (e.g. schizophrenia and/or bipolar disorder). PSZrel and PBDrel were excluded only if they had a medical condition that prevented participation.Participants provided written informed consent before participating in the study. The study protocol was approved and monitored by the MVAHCS and the University of Minnesota Institutional Review Board and adhered to the Declaration of Helsinki. Participants were administered the Structured Clinical Interview for the DSM-IV-TR Axis-I Disorders-Patient Edition  was then tested for a relationship to the visual behavioral task using Pearson's correlation against the parallel surround condition. The only measures that showed a correlation with surround suppression were visual acuity and estimated IQ (see p = 0.298).For each measure, potential group differences were assessed by 1-way ANOVA. Any measure that showed a potential group difference . Stimuli were generated using PsychoPy . The target and reference gratings were presented at a randomly selected orientation  on each trial; the orientation of the target and reference was the same on a given trial. The orientation of the surround was controlled relative to the target surround. There was also a “no surround” condition in which both sides of the screen appeared identical (i.e. only the target and the reference elements appeared).The contrast of the target stimulus was always 80%; the contrast of the reference stimulus was adjusted to achieve a match in perceived contrast. The side on which the reference stimulus was presented was randomized, so the target (plus surround) occurred on both sides of the screen with equal probability. Participants responded with a two-button button box to indicate whether the circular patch on the left side or the right side of the screen appeared to have higher contrast.For each condition, the contrast of the reference grating was controlled by a separate Psi staircase  implemenCatch trials were also embedded in the task . There were 48 catch trials, evenly divided between parallel and orthogonal surrounds and randomly assigned to the near or far condition. On a catch trial, the reference contrast was fixed at 30%. On these trials, participants should have always pressed the button that indicated that the non-reference side (target with surround) was of higher contrast. Performance on catch trials was used to assess participant engagement in the task and compliance with task instructions.One experimental run contained 48 staircase trials for each condition. Conditions were not blocked; trials from different conditions were mixed together because the task never changed. Participants completed one experimental run each, providing a single estimate of perceived contrast for each of the 11 conditions . The experimental run was paused four times so participants could rest their eyes and adjust their seat, verbally telling the experimenter when they were ready to continue.Data from each participant were analyzed if they met the following criteria: accuracy on catch trials was better than 75% and their behavior indicated that perceived contrast of the target with near surround at 0 degrees and 20 degrees relative orientation was reduced. This last criterion was in place to eliminate participants who could not selectively attend to the central targets and instead reported the overall (target plus surround) pattern. A total of 30 datasets were discarded because they did not meet these criteria , leaving a total of 138 datasets for analysis .For each of the 11 conditions , perceived contrast was calculated as the mean of the last 3 threshold estimates produced by the Psignifit staircase  for thatFollowing the a priori hypothesis that near and far surround suppression are mediated by different neural mechanisms, separate repeated measures ANCOVAs (rmANCOVAs) were performed to assess main effects of group and three of the surround conditions  and interaction between group and surround condition while controlling for visual acuity. Although we ultimately decided to include visual acuity as a covariate, such a decision is determined by theoretical perspective . Given the lack of certainty around this issue, we also report our main results without including acuity as a covariate . AdditioBayes factors were computed  represent our dependent variables of interest: the offset parameter (o) represents orientation-insensitive (i.e. untuned) gain-control while the magnitude (M), and tuning width (w) parameters jointly characterize orientation-dependent suppression. Fitting was done in Python using scipy.optimize.curvefit nonlinear least squares fitting algorithm . The model equation and parameters used to generate the simulated suppression tuning curves are fully detailed in the discussion. In the original model, attention provides multiplicative enhancement of neuronal responses to stimuli: focal attention (cA) enhances only the target response; distributed attention would enhance both target and surround responses. If this multiplicative modulatory term (cA) is instead used to represent the more general concept of “amplification following segmentation,” then either low acuity or broadly distributed attention (or a combination of the two) will result in reduced amplification of the center  and therefore stronger divisive normalization (response suppression) by the surround. Further, widely reported deficits in cortical untuned gain control associated with schizophrenia  = 3.59, p = 0.01, η² = 0.099; Fcondition = 164.51, p < 0.001, η² = 0.557; p = 0.08, η² = 0.053). A post hoc Bayes factor (BF) was computed to assess the lack of group differences observed for the 0 degrees condition while controlling for acuity by including acuity in the null hypothesis model. The 0 degrees BF was 0.001 (i.e. lack of group difference was 1000 times more likely). For comparison, the 90 degrees BF was 102.6 in favor of group differences.A repeated measures analysis of covariance was run on the three key near surround conditions , with Huynh-Feldt correction (ε = 0.991) and visual acuity as a covariate. This revealed significant main effects of group and condition (F−θ/w + oP = −Me) results for each group. Group differences in the three fit parameters were assessed with 1-way ANCOVAs, using acuity as a covariate. Previous work suggests that PSZ have broader orientation tuning (w parameters (broader orientation tuning), but there was not a significant difference between groups  = 1.06, p = 0.38, η² = 0.033). The M parameter showed a marginal effect of group that was not significant  = 2.19, p = 0.07, η² = 0.066) when acuity was entered as a covariate, but was significant without acuity as a covariate  = 4.95, p < 0.001, η² = 0.139). Follow-up pairwise comparisons showed that PSZ exhibited less negative o parameters when compared both to HCs and to BPDrel .n tuning , and PSZiate see . The untp = 0.02, η² = 0.090; Although there were no significant group differences in measured acuity see , acuity o parameter tracked meaningfully with individual differences in atypical sensory experiences using the SGI. To avoid the psychometric pitfalls of sum scores . Loadings greater than 0.3 for each factor are presented in p = 0.037). This third factor loaded most heavily on over-inclusion items such as “I notice background noises more than other people” and “I seem to hear the smallest details of sound” although it also loaded onto a few perceptual-modulation items  and fatigue/stress items (e.g. “When I'm tired sounds seem amplified”). For BPRS, five factors were extracted based on the most negative BIC value (χ²(166) = 342.48, p < 0.001, TLI = 0.71, CFI = 0.83, RMSEA = 0.09). None of the factor score estimates for these factors correlated meaningfully with individual differences in the offset parameter.Finally, we conducted exploratory correlational analyses to test whether untuned gain control as measured by the m scores , we condm scores . The fouo) for PSZ is consistent with previous reports of weakened untuned gain control associated with PSZ  were not significantly different between groups and we did not observe a significant interaction between group and condition. Having said this, post hoc Bayes factors provided strong evidence of a lack of difference between groups for the parallel-surround condition and strong evidence of group differences in the orthogonal condition. The preservation of suppression in the parallel-surround condition (i.e. lack of group difference) is surprising and unique to the specific configuration we used. Although we are unaware of this effect being observed previously in the context of surround suppression, analogous findings have been reported in the context of perceptual grouping tasks in which PSZ performed similar to controls when grouping cues were strongest  were excluded from analysis. It is noteworthy that approximately half of each of our experimental groups  had vision that was not corrected to normal (20/20) during the experiment. Our measurement of acuity was a simple Snellen eye chart at the 2-meter viewing distance that would be used for the task, with participants using any prescription lenses they had brought with them. It is possible that the relatively poor acuity across all groups was a consequence of the visual working distance being a poor match for the correction a given participant was using. To further complicate the matter, we did not observe significant differences in acuity between HC and PSZ as has been reported previously  does not fully account for the observed relationship observed between acuity and suppression across all participants: whereas low-acuity HCs demonstrated greater suppression at all orientations, low-acuity PSZ showed reduced suppression for orthogonal surrounds resulting in greater sensitivity to orientation. Thus, to explain the full pattern of data, we would need to posit that (1) all participants experience increased suppression when acuity reduces segmentation cues, and (2) of the PSZ group, only individuals with low acuity experience reduced untuned gain control (orientation-insensitive inhibition). Further work is necessary to determine the neural mechanisms underpinning this finding that acuity moderates the relationship between clinical group and suppression. The importance of acuity and retinal health in predicting schizophrenia has received increased attention recently, and rightly so . UnderstAlthough other researchers have found group differences in acuity with evidence that the difference is neural in origin , we did Within the framework of the normalization model discussed above, the intermediate performance of PSZrel could arise from a decrease in the semi-saturation term (σ) in the denominator used to regulate suppression  suppression mechanisms while evidence for group differences in orientation-dependent suppression mechanisms was mixed. Unexpectedly, the association between group and orientation-dependent suppression was moderated by acuity. Given recent findings of surround suppression deficits in a variety of psychological disorders including schizophrenia, bipolar disorder, autism and depression , this wo


Background: The prevalence of disabling conditions is increasing globally. Rehabilitation improves function and quality of life across many conditions, particularly when applied intensively. The limited workforce, however, cannot deliver evidence-based intensive rehabilitation. By providing individuals with the tools for self- rehabilitation, technology helps bridge the gap between evidence and practise. Few people, however, can access rehabilitation technology. Barriers such as cost, training, education, portability and poor design stand in the way of equitable access. Our group of engineers and researchers have established a centre dedicated to developing accessible technology through close, frequent engagement with users and industry.Methods: The centre employs a co-creation model, coupling engineering and science with user experience and industrial partnerships to develop accessible technology and associated processes. Due to the complexity and size of the challenge the initial focus is stroke. Recruited through a medical charity, participants, with a wide range of disabilities, use prototype and commercial technology during an 8-week rehabilitation programme with supervision from health professionals. The centre includes de-weighting systems, neurostimulation, virtual reality, treadmills, bespoke rehab games, communication apps, powered exercise equipment and gamified resistance equipment. Standard outcome measures  are recorded before, during, immediately after, and 3 months after the intervention and used in combination with an interview to design the initial rehabilitation programme, which is reviewed fortnightly. Qualitative methods (surveys and interviews) are used to capture personal experiences of the programme and individual technology and an advisory group of stroke survivors help interpret outcomes to feed into the technology design process. Ethical approval has been granted for a pilot cohort study with stroke survivors, which is currently underway (01/09/2021–31/12/2021) investigating acceptability and feasibility, due to report findings in 2022.Discussion: Through partnerships, research collaborations and a co-creation model a new centre dedicated to the development of accessible rehabilitation technology has been launched and currently undergoing acceptability and feasibility testing with stroke survivors. The centre, through its close engagement with users and industry, has the potential to transform the way rehabilitation technology is developed and help revolutionise the way rehabilitation is delivered. Since the industrial revolution improvements in disease prevention  have had a transformative effect on the health and longevity of most humans. This has been matched by cures for many diseases through surgical and drug interventions. Where disease and injury can't be cured there are well-developed nursing and medical procedures to maintain an individual's quality of life and palliative care to support the dying .The undoubted success of these health strategies has resulted in people living longer, healthier lives . The ageTechnology has long been suggested as a means to bridge the gap between evidence-based rehabilitation and what can realistically be provided by health services . While tIn this paper we detail the methods employed by the first engineering department (in Scotland) dedicated to using a co-creation framework for designing rehabilitation technology and outline the initial feasibility study. The intention of this approach is to produce technology that is useful; meaning both effective for recovery and accessible to all who need it.While the research team have a number of questions related to the specific effects of rehabilitation and optimisation of a technological approach to self-rehabilitation, the primary questions for the centre are initially concerned with acceptability and feasibility of the overall approach, including safety, adverse events, participant acceptability, recruitment, required levels of supervision, participant training and motivation.The guiding principle of the centre is co-creation through frequent and intensive user engagement. To enable this we have established a fully operational rehabilitation clinic equipped with commercially available, and prototype, rehabilitation technology, including de-weighting systems, non-invasive neurostimulation, virtual reality, treadmill, bespoke video games and tablet apps, powered exercise and gamified resistance equipment (see n = 7) has been designed to test the feasibility of this approach as well as participant acceptability. A mixed methods approach will be used to answer the key questions, including recruitment rates, retention rates, adverse events, programme compliance, level of assistance/supervision required, change in function and participant opinions on specific pieces of technology as well as the overall programme. Recruitment to the pilot commenced on 01/09/2021 and will end on 18/11/2021.A pilot cohort study with stroke survivors . Individuals subsequently contacting the centre are assessed for eligibility based on the criteria detailed in Once eligibility has been established an initial appointment is made to complete the informed consent process and let participants become familiar with the centre and some of the technology they may use. Participants, along with the research team, are also asked to adhere to the institutional Covid-19 restrictions during this period, including social distancing, mask wearing and carrying out two lateral flow tests per week.The 8-week, technology enabled, rehabilitation programme is structured around the broad principles of intensity, feedback, cognitive engagement and aerobic activity  to addreOutcome measures were selected based on their reported acceptability  in the research literature and common place use in the local health service. Details are provided in A structured rehabilitation programme will then be developed, co-operatively, by the rehabilitation professional and participant including agreed goals, frequency of attendance, duration and details of individual pieces of equipment to be used, see below for example.Total work time 80 min, with rest, as appropriate, in between activities.Improve general fitness, standing balance, lower limb strength, walking (symmetry and speed) and range of upper limb movement.Hand/wrist exercises following suggested guideline.1. Mirror therapy for hand (20 min)a. Seated cross trainer, 10 minb. Recumbent cycling 5 min.2. Powered exercise equipment3. Speech therapy app, picture and word matching, 10 mina. Balance games to encourage increased loading on weaker side, 5 minb. Walking in VR environment at comfortable walking speed (0.44 m/s) + 10%, 5 minc. Balance games to encourage increased loading on weaker side, 5 mind. Walking game that requires frequent adjustments to position, 5 min.e. Walking in VR environment at comfortable walking speed with 5 × 20 s periods of fast walking .4. Treadmill (25 min), see  for illu5. Functional squat 10 min—targeting symmetrical loading. 10 Kg load.Following completion of the 8-week programme the outcome measures are repeated. A plan to continue a modified version of the programme will be discussed. Finally, an independent researcher will arrange a time (within the following 2 weeks) for a one to one interview by telephone or free to download teleconferencing software. The interview will be open with the following questions given as a guideline;(1) How did you find the treadmill/bike/balance trainer/arm trainer/communication app?Was there anything you found particularly difficult to manage?Was there anything you found particularly easy to manage?(2) Technology based feedback on rehabilitation progressFor example:Did you find the progress information provided by the equipment useful?Were you able to access it easily enough?Did you need help understanding it?(3) EffectivenessFor example:Did you achieve your overall goal?Do you think there has been any change in your communication/mobility/balance/confidence/overall quality of life? Can you give a specific example to describe how this aspect has changed?Was there any particular piece of equipment that you think helped more?(4) Need for supervisionFor example:How important do you think having someone present was?Do you think you need a trained person present?Was there any equipment you feel you could manage without any help?(5) Potential for home useFor example:Would you consider using any of the equipment at home or at your local sports centre?Would you anticipate any difficulties using this equipment at home or your local sports centre?What support do you think you might need/like to use this equipment at home?(6) Future plansFor example:Now that you have completed the rehabilitation programme will you continue any activities/rehabilitation exercises?Do you have any plans to continue with rehabilitation?(7) Ideas for improvementFor example:Now that you have experienced rehabilitation using machines and equipment do you have any suggestions for improvement?Thinking about the specific equipment you used do you think we could improve it?Do you have any ideas of your own that might help people recover from stroke?How do you think we could improve the overall experience for you?The primary objective for the pilot study is to assess feasibility and acceptability. Data on compliance (sessions completed as percentage of planned), recruitment (people consenting as percentage of those volunteering) and rates of adverse events will be reported with descriptive statistics. Acceptability will be explored from a thematic analysis of the interview transcripts.In addition any change in the outcome measures (before and after the intervention) will be described with 95% confidence intervals  or interquartile range (if not).In this paper we have outlined the rationale for a co-creation centre in rehabilitation technology, provided some of the operational detail and outlined the methods for our pilot study designed to test feasibility and acceptability of the overall approach before we plan future research.Our pilot study is currently underway with seven stroke survivors participating. Once all participants have completed the programme a 6-week review period will be used to collate findings and implement any changes through discussion with our user group of stroke survivors. Once this has been completed, the project will move into a new phase with plans to recruit greater number of participants to our technology enabled rehabilitation programme.Based on outcomes from the pilot we expect to make changes to operational processes such as recruitment, outcome measures, induction and even the way the individual programmes are structured, however the very positive views expressed by participants to the media, so far (see quotes below for the first two participants), have encouraged us to continue planning future projects, including, but not limited to:New devices to improve hand function for more severely affected individualsThe use of machine learning to tailor individual rehab programmes on a continuous basisRecruitment of sub-acute stroke participants.#1 “I love the sessions, even though they go so quickly. I think the long-term plan is that this unit will act like a drop-in gym for those who need it, which would be fantastic. And if something like this could be available across the country, it would be even better. It would be a shame for people to miss out” .#2 “It was taking me 40 seconds to walk up down the gym and now I can do it in 12 seconds. That's just in the space of a couple of weeks,” .Despite mounting evidence on the positive effect of rehabilitation, recovery from stroke and other disabling conditions continues to be both disappointing and highly variable , 32 leadThe optimal “dose” of rehabilitation remains unknown , a more The original contributions presented in the study are included in the article/supplementary material, further inquiries can be directed to the corresponding author/s.The studies involving human participants were reviewed and approved by University of Strathclyde Ethics Committee. The patients/participants provided their written informed consent to participate in this study. Written informed consent was obtained from the individuals for the publication of any potentially identifiable images or data included in this article.All authors listed have made a substantial, direct, and intellectual contribution to the work and approved it for publication.The centre was part funded by the Sir Jules Thorn Charitable Trust.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article, or claim that may be made by its manufacturer, is not guaranteed or endorsed by the publisher.


The “kinetic triple factors” were calculated by thermodynamic analysis. The results show that the three Ea are 132.49, 116.36, and 118.24 kJ/mol, respectively; thus, the addition of H2SO4 and NaOH increased the thermal hazard of TBPTMH. In addition, the characteristic parameters  of its thermal decomposition were determined, and the control temperature  of TBPTMH under the action of acid-alkali were further received. This work is expected to provide some guidance for the safe storage, handling, production, and transportation of TBPTMH in the process industry.Tert-butyl peroxy-3,5,5-trimethylhexanoate (TBPTMH), a liquid ester organic peroxide, is commonly used as an initiator for polymerization reactions. During the production process, TBPTMH may be exposed to acids and alkali, which may have different effects on its thermal hazard, so it is necessary to carry out a study on the thermal hazard of TBPTMH mixed with acids and alkali. In this paper, the effects of H Organic peroxides are substances produced by replacing hydrogen atoms in hydrogen peroxide with organic groups such as alkyl, acyl, and aromatic groups ,2. It isTBPTMH is a liquid ester organic peroxide, and its structural formula is shown in 0) of TBPTMH+BPO was higher than that of TBPB+BPO, but the heat release was lower. Therefore, it is safer to apply TBPTMH as an initiator than TBPB in the production of molding compounds. Tseng et al. [3 (6 N), H3PO4 (6 N), and H2SO4 on MEKPO. The study showed that MEKPO is very sensitive to acid, especially HNO3 at 6 N. The addition of acid in both adiabatic and open environments decreased its T0. Liu [2SO4, NaOH, Na2SO3 on CHP, BPO, and DCPO using DSC, TAM III, and VSP2. The results showed that the risk of CHP, BPO, and DCPO was significantly increased by the addition of the above influences. You [3 on the thermal hazard of lauryl peroxide (LPO) and to calculate its TMR and SADT. It was found that LPO was very sensitive to inorganic acids, and its hazard increased with the increase in HNO3 concentration. Moreover, after the addition of HNO3, two exothermic peaks of LPO were detected, and explosive 1-nitrododecane was formed in the product, which increased the danger of LPO significantly.Recently, a number of studies have paid attention to TBPTMH and organic peroxides with acid-alkali. A brief overview of the literature is shown in g et al.  used VSPces. You  et al. uThe above studies show that the thermal hazard of TBPTMH is high and is increased by contact between organic peroxides and acids and bases, so it is essential to study the effect of acids and bases on the thermal hazard of TBPTMH.2SO4 and NaOH. DSC and adiabatic calorimetry (Phi-TEC II) were used to analyze their thermal hazards. The thermodynamic analysis was performed using thermodynamic theory to acquire the thermodynamic parameters, and the time to maximum rate under adiabatic conditions (TMRad), self-accelerated decomposition temperature (SADT), and reaction mechanism functions were further determined. This work investigates the effect of added acids-alkali on TBPTMH. Its decomposition in the adiabatic state and the mechanism function are also investigated, which is the innovative point of this paper. These findings are expected to provide guidance for the safe storage, transportation, and production of TBPTMH.In this paper, the effect of acid-alkali on the thermal hazard of TBPTMH was studied by adding H2SO4 and NaOH were obtained from Shanghai Lingfeng , and the purity of the above samples was higher than 98%. H2SO4 and NaOH were prepared at molar concentrations of 1 mol/L and 2 mol/L, respectively. The addition amounts of both influencing factors were 10%, and the upper oil phase was shaken by ultrasonication for 10 min and left to take for the experiment. The experimental flow chart is shown in TBPTMH used in this study was purchased from Wengjiang Reagent. HDifferential scanning calorimetry (DSC) is a technique that measures the variation in energy or power difference between the sample and the reference with temperature, which is controlled by a program ,11. It cIn this paper, the HP DSC 1 with low inertia and fast temperature rise and fall made by Mettler Toledo was used in experiments. The double safety of an explosion-proof sheet and sealing system made the experiment safer. The temperature range was 30–300 °C under a nitrogen environment with a flow rate of 50 mL/min. Of gold-plated crucible, 30 μL filled with 1.5 mg of sample was applied for each experiment.The characteristic parameters of a sample measured by DSC vary with the heating rate, so the results of DSC experiments can only be adopted for preliminary investigation of the thermal decomposition properties of the sample. The Phi-TEC II can detect the weak exothermic signal of the sample under adiabatic conditions to improve the accuracy of the results, which is one of the most commonly used apparatuses for thermal analysis. It can obtain the temperature and pressure curves with time during sample decomposition in an adiabatic environment ,14.In this paper, an adiabatic calorimetry (Phi-TEC II) manufactured by HLE was performed in the H-W-S (heat-wait-search) mode ,16. Phi-a), and reaction mechanism function) of a reaction by means of established mechanism functions [In chemical processes, thermodynamic analysis can be used as an important indicator parameter to determine optimal process conditions. In recent years, as the demand for chemical processes and the maturity of experimental techniques for thermal analysis have increased, this method has commonly been applied to evaluate the thermal stability of organics, inorganics, and polymers. Thermodynamic analysis is mainly used to obtain the “kinetic triple factors” , activation energy . Compared with the KAS and FWO methods, the activation energy calculated by it has higher accuracy, which is recommended by ICTAC and is widely used in fine chemical calculations [a is the activation energy, R is the gas constant, and T is the temperatureThe Starink method was deduced by analyzing the summary of the errors existing in the KAS, FWO, and Boswell methods. The Eulations ,21,22,23b and Ms are the mass of the sample ball and sample, respectively. Cvb and Cvs represent the specific heat capacity of the sample ball and sample.The ideal adiabatic environment is one in which the environment does not absorb the energy generated by the exotherm of the sample. However, in the actual Phi-TEC II experiments, the heat released by the sample is used to heat not only the sample itself, but also the ambient temperature of the cuvette. Therefore, Phi-TEC II does not fully satisfy the adiabatic condition either. The acquired adiabatic temperature rise and other parameters cannot be directly used to characterize the sample itself, and a thermal inertia factor must be introduced to correct the experimental data, as shown in Equation (2) ,25,26: :(3)lnk=−By choosing a different reaction order, the graph of lnk−1/T was plotted. Substituting the experimental data, a series of points at a different reaction order can be calculated. These points were fitted and analyzed, and the best-fitting line was selected, from which the activation energy and pre-exponential factor can be further calculated.2SO4 and NaOH, and to guide the determination of the mechanism function of the sample by the reaction level obtained.The Coats–Redfern method solves for the reaction activation energy by means of a mechanism function, which is usually used to probe the thermal hazard of a substance in depth, as shown in Equation (4) ,29,30,31ad is the time to maximum reaction rate under adiabatic conditions, which is also known as the adiabatic induction period, as shown in Equation (5) [ad can be used to describe the time to take protective measures when a decomposition reaction is runaway, which is usually considered to be proportional to the length of time to take protective measures. In chemical processes, we usually focus on the temperature corresponding to TMRad = 8 h and TMRad = 24 h.ad is the adiabatic temperature rise, C is the specific heat capacity of the sample, and Tf is the maximum decomposition temperature under adiabatic conditions.TMRtion (5) . TMRad cNR is the non-return temperature, M is the mass of the drug package, R is the gas constant, U is the heat transfer coefficient of the packaging material, and S is the surface area of the package.SADT is the lowest temperature at which self-accelerated decomposition of a chemical substance using packaging occurs ,34, whicDuring actual production, transportation, and storage, if the ambient temperature is higher than the value of SADT, the chemical will incur self-decomposition. The heat generated by the reaction is used to heat the reaction system, resulting in an increase in external temperature, which further promotes the decomposition of the chemical. Therefore, the ambient temperature of the chemical should be lower than the value of SADT, the temperature of the chemical should be recorded, and emergency rescue countermeasures should be formulated to prevent the reaction from occurring and causing more damage.2SO4 and NaOH are shown in Heat flow curves of pure TBPTMH and under the action of H0, Tp, and ΔH of TBPTMH thermal decomposition are 118.31 °C, 150.38 °C, and 687.42 J/g. Therefore, if TBPTMH decomposes and accumulates heat, a runaway reaction can easily occur.As can be seen in 2SO4 and NaOH remains as a single decomposition exothermic peak and does not change the decomposition peak of TBPTMH. According to the average of the data obtained from 0 of adding H2SO4 and NaOH are 112.54 and 117.86 °C, respectively. It can be seen that the addition of H2SO4 and NaOH reduced the T0 of TBPTMH and made TBPTMH more decomposable, with a peak temperature (Tp) of 148.88 and 149.74 °C and ΔH of 751.49 and 686.44 J/g. Compared with the ΔH of TBPTMH, it was found that the addition of acid-alkali made it higher, which indicated that more heat was generated, and the decomposition was more dangerous. Therefore, special attention should be paid to avoid contact with acid-alkali during the production, transportation, and storage of TBPTMH.As can be seen in 2SO4 and NaOH under adiabatic environment are illustrated in The temperature–time curves(a) and pressure–time curves(b) of TBPTMH and the addition of H0 after adding H2SO4 and NaOH is 81.58 and 83.2 °C, respectively. Compared with pure TBPTMH, their T0 was reduced, which was consistent with the conclusions of previous DSC experiments. ∆Tad rose particularly significantly with the addition of NaOH, close to about 20 °C, and the maximum temperature rise rate ((dT/dt)max)) and maximum pressure rise rates ((dP/dt)max)) were 192.85 °C/min and 168.36 bar/min, respectively, which should obviously indicate extra caution about the contact of NaOH with TBPTMH.In MATLAB software was used to fit the experimental data according to the least-squares method. To ensure the accuracy of the results, the DSC experimental data with a conversion (α) range of 10–90% and an interval of 10% each time were selected. Finally, the activation energy could be determined according to the slope of each fitting line, and the fitting results are outlined in The activation energies corresponding to different α are specified in 2SO4. In other words, the effect extent of the influencing factors is: NaOH < H2SO4.According to ad, dT/dtmax, and dP/dtmax of pure TBPTMH are all increased substantially, but T0 is decreased. This indicates that the decomposition of TBPTMH is more likely to occur under adiabatic conditions, and the decomposition rate is faster, which can lead to serious accidents in a short period of time.The adiabatic data were corrected according to Equation (2), and the corrected results are shown in 2SO4 and NaOH was in an ideal adiabatic environment, ΔH and ΔTad were increased in magnitude. In comparison, it was found that the two influencing factors of adding H2SO4 and NaOH could increase the thermal hazard of TBPTMH, especially for the NaOH. ΔTad was as high as 720.69 °C. dT/dtmax reached 813.83 °C/min after adding NaOH, which indicated that if the decomposition reaction of TBPTMH occurs after contacting with NaOH, the reaction will become violent, and the temperature and pressure will increase sharply, resulting in a short time to take safety measures, making evacuation and emergency more difficult to achieve.When the TBPTMH with the addition of HThe best-fitting straight line was selected by choosing a different reaction order (n) and fitting lnk-1/T, and the fitting results are disclosed in a for TBPTMH and added H2SO4 and NaOH were 129.98, 115.47, and 117.70 kJ/mol, respectively, and n were 2.6, 1, and 2.3, severally. The activation energies calculated by the Starink method were slightly lower than that in the adiabatic environment. This indicates that accidents are more likely to occur under adiabatic conditions than in open space. All the fitting coefficients are above 0.995, indicating that the results are accurate.Based on the fitting Equation (3), the adiabatic kinetic calculation results are enumerated in P-2SO4 and NaOH are displayed in According to Equation (4), a plot is made for 1/Ta of TBPTMH and the addition of H2SO4 and NaOH are 138.61, 113.40, and 118.72 kJ/mol, respectively, and the reaction orders are 1.16, 0.94, and 0.94, individually. Each n value is close to 1. The mechanism functions with n around 1 were selected from a was compared with that calculated by the Starink method. The mechanism function with high Ea and the great correlation coefficient was selected. Different G(α) from From a results in a calculated by the No. 16 mechanism function were the most similar. The difference of each Ea was < 10%, and the 10 correlation coefficients were all above 0.94, indicating the reliability of the results. The mechanism functions of TBPTMH and the addition of H2SO4 and NaOH are Comparing the Ead versus temperature was calculated according to Equation (5), as shown in A plot of TMRD8 (the temperature corresponding to TMRad = 8 h) was chosen in this paper to simulate the real operating temperature. The TD8 of TBPTMH is 63.71 °C, so the temperature should be controlled below 63.71 °C during the actual operation.According to the evaluation criteria of the possibility of accidents, TD8 with the addition of H2SO4 and NaOH are 59.16 °C and 60.51 °C, respectively. Therefore, the TD8 ranking of the three systems is: TBPTMH > TBPTMH + NaOH > TBPTMH + H2SO4. In other words, the effect content of the influencing factor is: NaOH < H2SO4. H2SO4 has the most significant effect on the decomposition of TBPTMH. Therefore, the contact hazard of TBPTMH with H2SO4 is greater compared to NaOH.The TIn this section, the SADT values for different packaging mass according to the Semenov model were determined, and the results are presented in From 2SO4 and NaOH added are also summarized in The SADT of the solution with H2SO4 are recommended to be 39.95 °C and 44.95 °C. The control temperature and the alarm temperature of TBPTMH under the action of NaOH are 39.73 °C and 44.73 °C, respectively. Therefore, to ensure safety during production, storage, and usage, the ambient temperature of TBPTMH and the addition of H2SO4 and NaOH should be controlled below 45, 40, and 40 °C, respectively.In general, the relationship between the control temperature and alarm temperature of the sample and SADT is shown in 2SO4 and NaOH on the thermal hazard of TBPTMH. The crucial conclusions are summarized as follows:To better guide the safe storage, transport, and production of TBPTMH, the DSC and Phi-TEC II were used to study the effect of adding H2SO4 and NaOH does not change the shape of the exothermic peak, but reduces the T0. In an adiabatic environment, the same conclusion can be obtained. The Starink method showed that the Ea of TBPTMH and the addition of H2SO4 and NaOH were 132.49, 116.36, and 118.24 kJ/mol, respectively, and the effect content of the influencing factor was NaOH<H2SO4. In an adiabatic environment, Ea were 129.98, 115.47, and 117.70 kJ/mol, respectively. The mechanism functions of the abovementioned systems were all No. 16: 2SO4 and NaOH are recommended to be lower than 45, 40, and 40 °C, respectively.The DSC experiments found that the addition of H


No particular complications were associated with both of our programs. We conclude that a hybrid rehabilitation program in low-risk ACS patients is feasible, safe, and as beneficial as a traditional program organised in a rehabilitation centre, at least in a short-term. A longitudinal follow-up should nevertheless be organised to examine the long-term impacts of this hybrid rehabilitation program.The aim of the study was to investigate the efficiency, the feasibility, and the safety of a hybrid cardiovascular rehabilitation program in low-risk acute coronary syndrome (ACS) patients. Sixty low-risk patients with stable clinical status who experienced an ACS in the previous 3 months were included in a 3-week rehabilitation program. The patients were randomized either to a group performing the rehabilitation totally in a rehabilitation centre or partially (only the first 5 days) and then in sport centres equipped for supervised adapted physical activities. The sport centres were located in the vicinity of the patient’s home. Both rehabilitation programs entailed endurance and resistance training and educational therapy. Before and after rehabilitation, cardiorespiratory functions were measured. Similar and significant improvements in peak  Survivors of a first acute coronary syndrome (ACS) deserve a specific care as the risk of experiencing a second cardiac event is significant . Centre-Nevertheless, the prescription of such rehabilitation in cardiac rehabilitation centres is rather low worldwide both in developed and less developed countries . This siOf course, this situation is unsatisfactory. Moreover, it has been observed that prescription rate is inversely correlated with patient risk. In other terms, rather low-risk patients are more easily assigned to cardiac centres than more high-risk patients are. This situation is unfortunate since it has been shown that in a 1-year survival study after acute myocardial infarction, the greatest mortality reduction was observed in high-risk patients who followed cardiac rehabilitation in a rehabilitation centre . CardiacIn the context of health prevention, treatment, and rehabilitation, a key issue is to provide the best available and most cost-effective intervention. Considering precision and personalized medicine, the importance of patient profiling to identify specific needs is crucial. Such an approach has advantages as individualized intervention can be tailored to specific outcomes according to the individual capacities. Therefore, it is logical to devise different kinds of procedures for low-risk and high-risk ACS patients. The high- or higher- risk patients should have priority for care in cardiac centres.The key issue for relevant care of low-risk ACS patients concerns the prevention of secondary cardiovascular disease through a multidisciplinary rehabilitation to reduce cardiovascular disease risk profile and improve exercise capacity. To achieve this purpose, organising the whole rehabilitation procedure in a cardiac rehabilitation centre may not be necessary. In the recent years, several alternatives have been explored. For instance, home-based or different kinds of hybrid cardiac rehabilitation have been evaluated and found to be of rather similar impact as traditional rehabilitation for different kinds of cardiac patients ,7. HowevTherefore, to increase access and participation to cardiac rehabilitation, to increase available options for prescribing cardiac rehabilitation, and to enhance long-term patient adherence to the program, we designed a hybrid cardiac rehabilitation model where the first stages of the rehabilitation are conducted in a traditional rehabilitation cardiac centre and the following in sport centres located in the vicinity of patient’s home under the supervision of trainers/educators who have specialised training in adapted physical activity. This study is investigated in terms of feasibility, efficiency, and safety.Sixty low-risk ACS patients with stable clinical status participated in two types of rehabilitation programs. The inclusion criteria were the onset of an ACS in the 3 previous months possibly treated with a percutaneous transluminal coronary angioplasty, with a low evolution risk profile , preserved systolic left ventricular function with an ejection fraction higher than 50%, the absence of any residual myocardial ischemia at rest and during exercise, no severe ventricular arrhythmias at rest and during exercise, and an adapted blood pressure rise during exercise. Moreover, patients were over 18 years old and affiliated to the French social security system . Exclusion criteria included locomotor disability, cognitive issues, or disabling disease that might interfere with the exercise protocol, all contra-indications to exercise test or training sessions, as well as abnormal symptoms during the rehabilitation process. Study protocols complied with the Helsinki declaration and was approved by the Ethics Committee of Biomedical Research . All of the subjects were informed about the study procedures and gave their written consent.The patients were referred for cardiac rehabilitation after an ACS and then randomized either to a group performing the training totally in a rehabilitation centre (3 weeks) or partially (only the first 5 days) and then during 3 weeks in sport centres equipped to supervise adapted physical activities until the end of the program (hybrid rehabilitation program). The sport centres were located in the vicinity of the patient’s home. Allocation to each group was determined by computer-generated random numbers. Demographic data were gathered before and after the traditional and the hybrid rehabilitation and included low-density (LDL) and high-density lipoprotein (HDL) cholesterol levels. Evaluations of patients were performed at the rehabilitation centre before and after both rehabilitation programs through a symptom-limited cardiopulmonary maximal exercise test and lower and upper limb strength tests .2), carbon dioxide production (2), expiratory flow (VE), and standard respiratory parameters were continuously monitored and averaged every 10 s. The first ventilatory threshold (VTh1) was evaluated using the Beaver and Wasserman method [The subjects performed a standardized symptom-limited cardiopulmonary maximal exercise test on an electrically braked stationary cycle ergometer  using a ramp protocol, before and after the rehabilitation programs as previously described . After an method .After a 10-min standardized warm-up, a one-repetition maximum strength test is performed according to Jidovtseff et al. . A similEach patient performed a set of 10 repetitions with a weight that was approximately 30–40% of the predicted 1RM. Patients rested for 3–5 min after which a set of 4–6 repetitions was completed with a load of approximately 50–70% of the predicted 1RM. After a 3–5 min rest period, the weight was increased to an estimated 90%RM load with a single repetition. The weight was then increased until failure was reached. The heaviest successfully lifted weight was considered as the 1RM. Again, there was a 3–5 min rest period between each attempt.In both rehabilitation programs, the patients were followed by a multidisciplinary team with cardiologists, medical doctors, specialized trainers in adapted physical activity, and dietician specialists. In both programs, the first five days were spent in the cardiac rehabilitation centre. Patients were included in the study groups, medically checked, informed about their condition and pathology through a therapeutic education program, given recommendations for their diets, and evaluated for physical prescription through exercise tolerance and strength tests. In both programs, the first training sessions were then performed at the cardiac rehabilitation centre for the first three days. The physical activity programs combined both endurance and resistance training and the prescription defined the training intensity of the exercises through cardiac frequency targets, the corresponding power output or exercise intensity on Borg scale.Two sessions of aerobic training were organised each day. One session consisted of exercises performed on a cycle ergometer as follows: a 5-min warm-up period followed either by 20 min of exercise at an individualized target intensity heart rate recorded at the first ventilatory threshold during the first exercise tolerance test and a 5-min period of active recovery or by 12 sequences of 30 s at 110–150% of the first ventilatory threshold workload, followed with 60 s at 40–80% of the first ventilatory threshold workload ,14. The In case of an abnormal cardiac event occurring during this initial phase in patients belonging to the hybrid program, the patients were excluded from the study. At the end of this first 3-day training, an agreement contract containing individualized objectives was signed by the patient. A booklet presenting the sessions that the patients would follow in the second part of the program was also given with individualized intensity training targets. Patients or the sports centre personal trainer were asked to fill in description, duration, and intensities of each session right after each session. They were also asked to report any adverse events.Concerning the second part of the rehabilitation program, the groups of patients continued their protocols at either the cardiac centre or in a delocalized sport centre. The patients who stayed at the cardiac centre continued the same program as described above, 5 times per week. Those who went to the sport centres were asked to follow a 3-week program with 3 sessions of 1 h per week. The session consisted of 30–40 min of aerobic exercise performed on a cycle ergometer similar to the training performed in the cardiac rehabilitation centre. The following 20–30 min were devoted to at least four to seven resistance training exercises performed in a similar fashion as the training received in the cardiac rehabilitation centre. Besides the planned session at the sport centre, the patients were also asked to go for a 1 h walk each day.The first session at the sport centre was considered as an introductory session and was conducted by the patient personal trainer under the supervision of a member of the cardiac centre involved in the project. The following sessions were then conducted under the supervision of the patient personal trainer where the patient was continuously followed. Only one person was being supervised at a time.At the end of the hybrid rehabilitation program, a satisfaction score was given by the patients on scale from 0 to 10 (0 being the worst and 10 the best). Patients received a booklet proposing exercises  that were recommended to be performed on daily basis.Each patient’s personal GP was informed of his patient’s inclusion in the sport centre rehabilitation program and his collaboration was needed to ensure the quality of the rehabilitation follow-up care in case of a medical problem. Systematically, a patient appointment was scheduled with his GP during the 2nd week of the delocalized rehabilitation phase. The GP also had a direct phone access to the cardiologists and/or physical therapists of the cardiac rehabilitation centre as well as to the referred personal from the sport centre.Before addressing participants to a sport centre, the cardiac rehabilitation centre reviewed the sport centres in its region or district. The selected sport centres were requested to have professional educators in physical adapted activities or physical trainers, established protocol in case of emergency, and an automated external defibrillator available in the room where the training sessions were organised. In order to enhance the personnel abilities to supervise low risk ACS patient exercise, a specific in-service training was organised by the cardiac centre. The training for the sports centre trainers focused on gauging and developing knowledge in cardiovascular diseases, the specific needs of low-risk ACS patients, monitoring of training, and emergency protocols in case of health problems during the training sessions.2 peak comprised between 3 to 5 mL/kg/min with a non-inferiority of 3 mL/kg/min), a one-sided type I error of 0.025 and 80% power, a total sample size of 42 patients (21 assigned to each group) is required. This number of recruited patients was increased to 30 in each group in order to prevent possible dropouts.To assure a non-inferiority hypothesis on the primary outcome . Kolmogorov–Smirnov and Lilliefors tests were used to check the normality of the distribution. Student’s The characteristics of the patients who completed the entire study are presented in p < 0.05) in 2 and power output peaks as well as in peak heart rate and power output at the first ventilatory threshold were observed. No significant changes in upper and lower limb strength were noted. The patient serum HDL-cholesterol concentration stayed stable in both groups when comparing the data obtained before vs. after the rehabilitation . Concerning the changes in LDL-cholesterol, we observed a significant decrease in both groups when comparing the data obtained before vs. after both rehabilitations . However, the changes were considered similar in both groups as no significant interaction was found. No adverse events were noted during the hybrid rehabilitation program. All the participants from this program performed the 1 + 9 scheduled sessions that were planned. Those patients also gave a satisfaction score of 9.9/10 for their rehabilitation. From the 28 participants belonging to the hybrid program, 18 of them continued regular visits to the sport centre after the rehabilitation program officially ended.The main changes observed in both programs are reported in In this preliminary trial, we developed a concept of a new hybrid cardiovascular rehabilitation program where the cardiac rehabilitation centre is kept as the control tower of the whole rehabilitation processes but where a part of the rehabilitation is performed outside of its walls . This moThis model offers several potential advantages and could, at the same time, lower the medical cost dedicated to traditional cardiac rehabilitation. The savings could then be re-invested for the treatment of more severe ACS patients and allow more ACS patients access to cardiac rehabilitation programs after a cardiac event. This hybrid model could also facilitate the everyday life of low-risk ACS patients who are unable to stay, visit, or reach rehabilitation centres for various reasons; increase the amount of physical activity in patients; improve the patient satisfaction and adhesion to the proposed rehabilitation program; help patients incorporate sports centre visits into daily life; help patients incorporate long-term healthy lifestyle changes concerning regular physical activity; and improve the quality of life in patients.2 and power output peaks were similar between this hybrid model and regular rehabilitation and consistent with the outcomes published in the scientific literature [In this time of precision and personalized medicine, such a tailored approach needs to be first investigated in terms of feasibility, efficiency, and safety. The main results of this investigation are that it is possible to safely organize such a rehabilitation program since we did not observe any special complications during the length of the study, and it seems to also be as effective as a traditional rehabilitation, at least in a short-term. We found that the changes in terature ,19. ConcPatient adhesion to the hybrid program was also high as no patients dropped out of the study in the second part of the program and all patients participated in every planned rehabilitation session. Another indicator of patient adhesion is that 18 of the 28 participants in the hybrid rehabilitation group continued to visit the sport facilities on regular basis after the study was completed.There have been many initiatives around the world to study alternatives to traditional cardiac rehabilitation in specialized rehabilitation centres. Such hybrid cardiac rehabilitations offer a mix of direct supervised centre-based and home-based cardiac rehabilitation with the use of special communication devices  between patients and health/medical supports. A recent meta-analysis concluded that hybrid cardiac rehabilitation globally showed similar efficacy to the traditional model, at least in the short term. However, the long-term outcomes are for the time being less clear and still need to be investigated ,20,21. 2 peak was observed in patients after hybrid rehabilitation compared to those following the traditional one [Some recent cost-efficient care delivery strategies using hybrid rehabilitation claimed to induce long-term health benefits. However, even though such long-term health benefits were shown, the authors nevertheless expressed that a significant stronger decline in onal one . Howeveronal one ,9. Our proposed individual tailored intervention focused on supervised adapted physical activity performed in specific sport centres in the vicinity of the patients’ living places. Such intervention is characterised by a reduced fee for medical supervision, is more practical for the patient who can more easily combine the rehabilitation with his/her own agenda. This aspect may help for program adhesion and long-term behavioural change. The proposed hybrid intervention is also in line with the recent possibilities of medically prescribed physical activities for chronic disease patients. This approach is rather new, and the patient’s service offers need to be developed both in France and in other parts of the world. Our hybrid intervention model could serve such a purpose and could even be generalised to any kind of low-risk patients . In rehabilitation centres, low-risk patients could be included on Mondays, instructed, and trained in the physical exercise programs and to the different components of the traditional rehabilitation. Then, if no difficulties or medical problems occur, they could be discharged on Fridays to pursue their rehabilitation using this hybrid model. A follow-up check-up visit could be scheduled at the traditional rehabilitation centre to measure progress after the program and/or at distance of the program.In conclusion, a new hybrid cardiac rehabilitation program in low-risk ACS patients —where the first part of the rehabilitation is organised in a cardiac rehabilitation centre and then in habilitated sport centres located near the patient’s home—is feasible, safe, and as beneficial as a traditional program totally organised in a rehabilitation centre, at least in a short-term. However, the present investigation is still at its preliminary phase, and the obtained results need to be confirmed with a larger number of patients and with a precise follow-up to investigate the long-term outcomes of such approach.


D-value of 5.9 µM as well as IC50 values of 4.0 µM in PBS and 10.2 µM in saliva. We thus introduce a rapid test for kynurenine as a potential biomarker for kidney transplant failure.Kynurenine is a tryptophan metabolite linked to several inflammatory processes including transplant failure, a significant challenge in transplant medicine. The detection of small molecules such as kynurenine, however, is often complex and time consuming. Herein, we report the successful synthesis of a fluorescently labelled kynurenine derivative, showing proper fluorescence and anti-kynurenine antibody binding behavior in a magnetic bead immunoassay (MIA). The fluorescent kynurenine–rhodamine B conjugate shows a K Kynurenine is a metabolite of the tryptophan degradation pathway linked to several inflammatory, metabolic, oncogenic  as well The link between inflammatory response and elevated kynurenine levels in serum via upregulation of IDO by pro-inflammatory factors renders kynurenine an interesting biomarker for clinically relevant inflammatory processes. Upregulated IDO and, therefore, elevated kynurenine levels of 3.9 ± 2.1 µM in serum have been found in the case of chronic kidney disease (CKD) in patients in the pre-dialysis stage. Values increase along with CKD severity up to 5.6 ± 2.3 µM because of chronic inflammation during CKD progression ,20,21,22Apart from transplant failure, elevated kynurenine levels have also been linked to titanium dental implants and to bone-osseointegration processes , as wellMost approaches in clinical trials to quantifying kynurenine from different tissues are focused on LC-MS ,34,35 orTo date and due to the small size of the kynurenine molecule, no fluorescently labelled kynurenine derivatives bound to an anti-kynurenine antibody are described in the literature. Likewise, there is no functional bioassay based on antibody binding of kynurenine described in the literature. Herein, we report the synthesis of a fluorescent, rhodamine B-labelled kynurenine derivative and demonstrate its binding ability to an anti-kynurenine antibody. We demonstrate that this denotes a promising approach for the development of a competitive kynurenine antibody-binding assay and ultimately a kynurenine rapid test from saliva and possibly other body fluids.4 in 1N NaOH. Flash column chromatography (FC) was performed on M&N Silica Gel 60 (0.063–0.200 mm). 1H NMR and 13C NMR spectra were recorded on a Bruker Avance I 200 (200 MHz), Bruker Avance II 400 , or Varian Unity 500 (500 MHz) spectrometer . Chemical shifts are reported in parts per million relative to solvent signals . Signals were assigned by first-order analysis and assignments were supported where feasible by 2-dimensional 1H, 1H and 1H, 13C correlation spectroscopy. Coupling constants are reported in hertz. UV/vis spectra were recorded on a PerkinElmer Lambda XLS+ UV/vis spectrometer  in a 10.00 mm QS quartz cuvette. Fluorescence was recorded on a Tecan Infinite M200 multiplate reader in commercially available 96-well multitier plates. Chemicals and reagents were purchased from Acros Organics, Alfa Aesar, Sigma-Aldrich, Carl Roth, Carbolution or ABCR and were used without further purification.TLC was carried out on Silica Gel 60 F254  with detection by UV light (254 nm) or by charring with 1% KMnO2Cl2 (250 mL) under N2 atmosphere. EDC  and DMAP  were added, and the mixture obtained was stirred for 15 min at room temperature. After the addition of propargyl alcohol  and after stirring overnight at room temperature, the mixture was washed 2x with 1M HCl (250 mL) and 1x with brine (250 mL). Drying the organic layer over Na2SO4 and evaporating the solvent yielded the crude product. Purification via flash column chromatography (CH2Cl2: MeOH 9:1) gave the final product: violet powder .Rhodamine B  was dissolved in dry CH1H-NMR : 8.66 , 7.88 , 7.78 , 7.38 , 7.09 , 6.95 , 6.88 , 4.65 , 3.68 , 2.46 , 1.36 .13C-NMR : 164.4 (C(=O)O), 157.6 (Ar-C), 155.6 (Ar-C), 134.0 (Ar-C), 133.8 (Ar-C), 131.3 (Ar-C), 131.3 (Ar-C), 131.0 (Ar-C), 129.3 (Ar-C), 115.1 (Ar-C), 113.4 (Ar-C), 107.4 (Ar-C), 96.4 (Ar-C), 78.5 (C), 77.8 (CH), 53.3 (CH2), 45.8 (CH2), 12.9 (CH3).2 protection of kynurenine was carried out by Boc-chemistry following Schotten–Baumann conditions: kynurenine  was dissolved in a water/THF 1:1 mixture containing NaOH . After the solution turned clear, Boc2O  was added dropwise at 0 °C, and the reaction was monitored by TLC. Complete conversion was observed after 90 min. Acidification using 10% HCl, followed by 3 extractions with ethyl acetate, drying of the organic layer and evaporation of the solvent yielded the crude product: a yellowish oil. Column chromatography (ethyl acetate) yielded the pure product: a yellowish powder .NH1H-NMR : 7.73 , 7.34 , 6.83–6.72 , 5.64 , 4.67 , 3.80–3.69 , 3.53 , 1.45 .13C-NMR : 199.54 (Ar-C(=O)), 172.43 (C(C=O)OH), 155.68 (Boc-C(=O)), 149.45 (Ar-C), 134.96 (Ar-C), 131.08 (Ar-C), 125.83 (Ar-C), 118.10 (Ar-C), 116.73 (Ar-C), 69.98 (Boc-tert-C), 52.60 (CH), 41.47 (CH2), 28.34 (CH3).1  and the corresponding azido-linker  were dissolved in CH2Cl2 (50 mL). EDC  and DMAP  were added, and the mixture was stirred at room temperature for 48 h. Washing 3× with 2 M NaOH (80 mL) followed by washing with brine (80 mL), drying over Na2SO4 and evaporation yielded the crude product. Column chromatography (ethyl acetate) yielded the pure product: an orange solid, which was used directly in the next step without further analysis .Boc-protected kynurenine 3  and propargyl rhodamine B 1  were dissolved in 30 mL of a mixture of CH2Cl2/MeOH/H2O 10:10:3. After adding an aqueous CuSO4 solution , TBTA  and Na ascorbate , the mixture was heated to 60 °C for 16 h. After cooling down, 20 mL of ddH2O was added and the mixture was extracted 3 times with 50 mL CH2Cl2, followed by drying over Na2SO4 and evaporation of the solvent. The pure product was obtained after column chromatography (CH2Cl2/MeOH 3:1) as a pink powder .Azido functionalized Boc-L-Kynurenine 1H-NMR : 8.20 , 7.91–7.85 , 7.83–7.79 , 7.47 , 7.08–7.00 , 6.97–6.90 , 5.04 , 4.65 , 4.44 , 3.74 , 3.63 , 3.48–3.45 , 3.44–3.40 , 3.30 , 3.00 , 1.33 , 1.20 .13C-NMR : 198.9 (Ar-C(=O), 172.1 (C(=O)-NH), 164.9 (C(=O)-O), 157.8 (Ar-H), 157.5 (Boc-C(=O)), 156.0 (Ar-C), 155.6 (Ar-C), 155.5 (Ar-C), 141.04 (Triazol-Ar-C), 134.6 (Ar-C), 133.9 (Ar-C), 133.8 (Ar-C), 133.7 (Ar-C), 131.3 (Ar-C), 131.2 (Ar-C), 131.0 (Ar-C), 130.9 (Ar-C), 129.6 (Ar-C), 129.2 (Ar-C), 125.3 (Triazol-Ar-C), 115.0 (Ar-C), 114.9 (Ar-C), 114.8 (Ar-C), 113.3 (Ar-C), 96.3 (Ar-C), 78.4 (tert.-C), 78.0 (NH(Boc)-CH), 77.8 (O-CH2), 70.1 (O-CH2), 70.0 (O-CH2), 69.9 (O-CH2), 69.6 (O-CH2), 69.3 (O-CH2), 68.0 (O-CH2), 58.6 (Ar-CH2), 53.2 (RhB-CH2), 49.8 (Ar-CH2), 45.8 (N-CH2), 28.7 (Boc-CH3), 12.9 (RhB-CH3).4 was carried out by dissolving conjugate 4 in 6 mL of CH2Cl2 containing 25% trifluoroacetic acid. After stirring at room temperature for 1 h, the solution was precipitated in 50 mL ice cold Et2O and centrifuged for 5 min at 4 °C and max speed. The precipitate was dissolved in ddH2O and lyophilized. Purification by HPLC yielded the pure product: a pink solid .Boc-deprotection of rhodamine B–kynurenin conjugate 1H-NMR : 8.20 , 8.11 NH), 7.93–7.84 , 7.85–7.78 , 7.47 , 7.05–6.99 , 6.97–6.89 , 5.05 , 4.47–4.41 , 3.75 , 3.63 , 3.54 , 3.50–3.42 , 2.92 , 1.20 .13C-NMR : 197.3 (Ar-C(=O)), 168.8 (C(=O)-NH), 164.9 (C(=O)-O), 158.2 (Ar-C), 157.9 (Ar-C), 157.5 (Ar-C), 155.5 (Ar-C), 141.1 (Triazol-Ar-C), 133.2 (Ar-C), 133.7 (Ar-C), 131.2 (Ar-C), 130.9 (Ar-C), 129.6 (Ar-C), 125.3 (Triazol-Ar-C), 117.5 (Ar-C), 114.9 (Ar-C), 113.3 (Ar-C), 96.3 (Ar-C), 70.1 (O-CH2), 70.0 (O-CH2), 69.9 (O-CH2), 69.0 (O-CH2), 67.1 (O-CH2), 58.6 (Ar-CH2), 49.8 (Ar-CH2), 45.7 N-CH2), 12.87 (CH3).1  and azidotetraethylene glycol  was added, TBTA  and Na ascorbate  were added, and the mixture was heated to 60 °C. The reaction was monitored by TLC. After 16 h, the mixture was left to cool down, followed by the addition of 25 mL of ddH2O. Extraction with 50 mL of CH2Cl2 3 times, followed by drying over Na2SO4, gave the crude product. Column chromatography (CH2Cl2/MeOH 3:1) gave the pure product: a pink oil .Propargyl rhodamine B terature ) were di1H-NMR : 8.22 , 7.89 , 7.82 , 7.48 , 7.04 , 6.96 , 6.95–6.92 , 5.06 , 4.56 , 4.45 , 3.76 , 3.65 , 3.49 , 3.46–3.41 , 3.37–3.34 , 1.22 .13C-NMR : 164.88 (C(=O)), 157.88 (Ar-H), 157.53 (Ar-H), 155.53 (Ar-H), 141.05 (Ar-H), 133.71 (Ar-H), 131.24 (Ar-H), 130.93 (Ar-H), 129.67 (Ar-H), 125.36 (Ar-H), 114.94 (Ar-H), 113.32 (Ar-H), 96.29 (Ar-H), 72.75 (CH2), 70.21 (CH2), 70.17 (CH2), 70.05 (CH2), 69.97 (CH2), 69.01 (CH2), 66.81 (CH2), 60.63 (CH2), 58.63 (CH2), 49.77 (CH2), 45.76 (CH2), 12.89 (CH3).®12-biotin, Sigma Aldrich, Taufkirchen, Germany) was added to 100 µL of an anti-kynurenine antibody . The mixture was incubated for 50 min at room temperature with gentle shaking. Excess Biotin-NHS was removed using a VivaSpin 500 centrifugal concentrator with 10 kDa MWCO (Sigma Aldrich).A total of 1 µL of a 6 mg/mL Biotin-NHS solution  was diluted to 1 mg/mL with 450 µL of PBS and pelleted on a magnetic rack for 2 min. The supernatant was discarded, and the beads were washed three times with 500 µL PBS. After the last washing step, 20 µL of the supernatant was replaced with 20 µL biotinylated antibody (0.5 mg/mL). The mixture was incubated for 30 min at RT under gentle shaking. The reaction was blocked by washing the beads three times with PBS containing 1.5% BSA and 0.5% Tween-20. The final concentration was 20 µg of antibody per 1 mg of beads.5 or rhodamine B-PEG-Linker 6 was incubated with 50 µL of antibody-conjugated magnetic beads and 50 µL of PBS (1:3 dilution). The beads were pelleted on a magnetic rack, and the unbound rhodamine B in the supernatant was quantified in a 96-well-plate with a fluorescence measurement of 100 µL of supernatant at 561 nm excitation and 592 nm emission, using a Tecan Infinite M200 multiplate reader. A standard curve of rhodamine B fluorescence intensity between 0 and 100 µM diluted 1:3 in PBS was used for the calculation of bead-bound conjugate 5 or 6.For 3 h at RT, 0–100 µM rhodamine B–kynurenine conjugate 5 and 50 µL of spiked PBS or artificial saliva  containing 0–250 µM native L-kynurenine for 3 h at RT on a hula shaker (1:3 dilution). Beads were pelleted on a magnet and fluorescence intensity of unbound rhodamine B–kynurenine conjugate 5 in 100 µL supernatant was measured in a Tecan Infinite M200 multiplate reader at 561 nm excitation and 592 nm emission. Bead-bound L-kynurenine was indirectly calculated by calculating the amount of displaced rhodamine B–kynurenine conjugate 5. Therefore, the fluorescence intensity of bound rhodamine B–kynurenine conjugate 5 without L-kynurenine was subtracted from the fluorescence intensity of samples with different L-kynurenine concentrations.Fifty microliters of antibody coupled beads was incubated with 50 µL of 12 µM rhodamine B-kynurenine conjugate 1 using Steglich esterification by EDC and DMAP, which was used as the fluorescent probe for click-conjugates 5 and 6.Chemical synthesis of the fluorescent L-kynurenine–rhodamine B conjugate was carried out as shown in 3, Boc-protection of the primary amine of kynurenine was carried out to obtain product 2, followed by attaching an amino-azido-tetraethylene glycol linker, synthesized according to a published protocol [3. Click reaction of 3 with 1 gave product 4, followed by Boc-deprotection to the fluorescent kynurenine probe 5. To study the effect of the linker as well as the rhodamine B moiety on antibody binding, rhodamine B clickamer 6 containing only a tetraethylene glycol linker [4.For the azido tetraethylene glycol L-kynurenine derivative protocol  yieldingl linker  was prep2O (cf. 5 and 6, containing a benzoic ester instead of a free benzoic acid at the rhodamine’s benzoic acid residue, is slightly shifted to 560 nm. Emission maxima were determined to be at 586 nm for the L-Kyn-4EG-RhB probe 5, while the 4EG-RhB clickamer 6 shows an emission maximum of 584 nm, indicating suitable absorption and emission properties for immunoassays.To determine emission and absorption maxima for the synthesized rhodamine B click-conjugates, UV/vis spectra of the compounds were recorded in ddH2O cf. . The abs5 and 6 as well as their competitive binding capability by using magnetic bead-bound anti-kynurenine antibodies, followed by incubation with the fluorescent conjugates and the subsequent fluorescence measurements of unbound conjugate in the supernatant. The tetraethylene glycol rhodamine B clickamer 6 was used as a negative control to exclude unspecific interactions between the rhodamine B or the tetraethylene glycol linker with the antibody. Antibody binding data are shown in 5 and native L-kynurenine.To determine the antibody binding capabilities of the synthesized fluorescent L-kynurenine conjugate, we elucidated the antibody binding capability of fluorescent conjugates 5, followed by a pelleting of the beads and the measurement of the fluorescence in the supernatant. The binding curves of 5 and 6 as well as the competitive binding curve are shown in To this end, antibodies were bound to magnetic beads in 1 µm diameter, followed by incubation with the target compounds, pelleted via magnet and the fluorescence measured in the supernatant. In the competitive assay, 0–83 µM L-kynurenine was incubated together with either 4 µM RhB-4EG-L-Kyn conjugate 5, the measured fluorescence in the supernatant decreases since more fluorescent conjugates bind to the antibody. 5 and 4EG-RhB clickamer 6 with and without incubation with the bead-bound antibodies. In higher concentrations, the 4EG-RhB clickamer 6 binding affinity also increases, probably due to unspecific interactions between either the linker or the rhodamine B residue. Apart from this, the difference in measured fluorescence for the L-kynurenine conjugate 5 is significantly stronger, showing a KD-value of 5.9 µM, and therefore, unspecific interactions between the 4EG-RhB clickamer 6 and the magnetic bead-bound antibody can be neglected.With increasing concentration of the fluorescent L-kynurenine conjugate 50 values of this competition were calculated to be 4.0 µM in PBS and 10.2 µM in saliva. As expected, the IC50 value in saliva is higher than in PBS due to interfering components such as enzymes and proteins in saliva samples.As shown in D-value of 5.9 µM for the L-kynurenine conjugate 5 and IC50 values of 4.0 µM in PBS and 10.2 µM in saliva for the competitive assay. Since an increase in kynurenine levels in saliva to 4.6 ± 1.6 µM under pathological conditions, compared to 0.7 ± 0.4 µM in the healthy subject, are observed [We successfully synthesized a fluorescent kynurenine conjugate based on rhodamine B with unique antibody binding properties, a promising component of a future rapid diagnostic test for kynurenine, a metabolite with a clinically relevant marker for the diagnosis of distinct diseases. Spectral properties of the products were elucidated, showing only a slight shift of 6 nm in the fluorescence emission maximum when compared to native rhodamine B. The antibody binding was investigated, and the magnetic bead assay showed a good sensitivity with a Kobserved , it thusobserved , even th