Correction: Development of an optimized AAV2/5 gene therapy vector for Leber congenital amaurosis owing to defects in RPE65.

The authors originally published this article under the incorrect license type; this has now been corrected and is published under the CC-BY license.

Development of an optimized AAV2/5 gene therapy vector for Leber congenital amaurosis owing to defects in RPE65.

Leber congenital amaurosis is a group of inherited retinal dystrophies that cause severe sight impairment in childhood; RPE65-deficiency causes impaired rod photoreceptor function from birth and progressive impairment of cone photoreceptor function associated with retinal degeneration. In animal models of RPE65 deficiency, subretinal injection of recombinant adeno-associated virus (AAV) 2/2 vectors carrying RPE65 cDNA improves rod photoreceptor function, and intervention at an early stage of disease provides sustained benefit by protecting cone photoreceptors against retinal degeneration. In affected humans, administration of these vectors has resulted to date in relatively modest improvements in photoreceptor function, even when retinal degeneration is comparatively mild, and the duration of benefit is limited by progressive retinal degeneration. We conclude that the demand for RPE65 in humans is not fully met by current vectors, and predict that a more powerful vector will provide more durable benefit. With this aim we have modified the original AAV2/2 vector to generate AAV2/5-OPTIRPE65. The new configuration consists of an AAV vector serotype 5 carrying an optimized hRPE65 promoter and a codon-optimized hRPE65 gene. In mice, AAV2/5-OPTIRPE65 is at least 300-fold more potent than our original AAV2/2 vector.

Statement of the Spanish Interdisciplinary Cardiovascular Prevention Committee (CEIPC for its Spanish acronym) on the 2012 European Cardiovascular Prevention Guidelines.

Based on the two main frameworks for evaluating scientific evidence (SEC and GRADE) European cardiovascular prevention guidelines recommend interventions across all life stages using a combination of population-based and high-risk strategies with diet as the cornerstone of prevention. The evaluation of cardiovascular risk (CVR) incorporates HDL levels and psychosocial factors, a very high risk category, and the concept of age-risk. They also recommend cognitive-behavioural methods (e.g., motivational interviewing, psychological interventions) led by health professionals and with the participation of the patient's family, to counterbalance psychosocial stress and reduce CVR through the institution of positive habits such as a healthy diet, physical activity, smoking cessation, and adherence to treatment. Additionally, public health interventions - such as smoking ban in public areas or the elimination of trans fatty acids from the food chain - are also essential. Other innovations include abandoning antiplatelet therapy in primary prevention and the recommendation of maintaining blood pressure within the 130-139/80-85 mmHg range in diabetic patients and individuals with high CVR. Finally, due to the significant impact on patient progress and medical costs, special emphasis is given to the low therapeutic adherence levels observed. In sum, improving cardiovascular prevention requires a true partnership among the political class, public administrations, scientific and professional associations, health foundations, consumer associations, patients and their families. Such partnership would promote population-based and individual strategies by taking advantage of the broad spectrum of scientific evidence available, from clinical trials to observational studies and mathematical models to evaluate population-based interventions, including cost-effectiveness analyses.

Ovine ear model for fully endoscopic stapedectomy training.

Endoscopic surgery of the middle ear is progressively gaining the interest of otologists, as technological advances have overcome some of its main drawbacks. The long learning curve required to master this technique, urges the search for models to practice it. After the validation of sheep's ear as a proper training model for microscopic stapedectomy, our objective is to demonstrate its adequacy for practicing stapes surgery but performed through a fully endoscopic approach. Endoscopic stapedectomy was performed by two surgeons in 40 sheep ears (20 specimens each). To analyze the effects of the learning curve on surgical success, complication rates and surgical time reduction, the sample was divided in two groups: group 1 being the first ten procedures of each surgeon, and group 2 the second set of stapedectomies. The impact of the operated side and the resection of the chordal spine were also studied. No statistically significant differences were found considering the operated side. A statistically significant improvement in some of the surgical steps was demonstrated comparing both groups and also after the resection of the chordal spine. Mean surgical time declined from 38 to 31.5 min (p < 0.05). Using this model for endoscopic stapedectomy, a learning curve was objectively demonstrated, along with other subjective appreciations such as improvement in depth perception and one-hand instrument handling. We believe that sheep ear is an optimal model for endoscopic middle ear surgery, as it allows for the acquisition of the skills required to master this technique.

Effective transplantation of photoreceptor precursor cells selected via cell surface antigen expression.

Retinal degenerative diseases are a major cause of untreatable blindness. Stem cell therapy to replace lost photoreceptors represents a feasible future treatment. We previously demonstrated that postmitotic photoreceptor precursors expressing an NrlGFP transgene integrate into the diseased retina and restore some light sensitivity. As genetic modification of precursor cells derived from stem cell cultures is not desirable for therapy, we have tested cell selection strategies using fluorochrome-conjugated antibodies recognizing cell surface antigens to sort photoreceptor precursors. Microarray analysis of postnatal NrlGFP-expressing precursors identified four candidate genes encoding cell surface antigens (Nt5e, Prom1, Podxl, and Cd24a). To test the feasibility of using donor cells isolated using cell surface markers for retinal therapy, cells selected from developing retinae by fluorescence-activated cell sorting based on Cd24a expression (using CD24 antibody) and/or Nt5e expression (using CD73 antibody) were transplanted into the wild-type or Crb1(rd8/rd8) or Prph2(rd2/rd2) mouse eye. The CD73/CD24-sorted cells migrated into the outer nuclear layer, acquired the morphology of mature photoreceptors and expressed outer segment markers. They showed an 18-fold higher integration efficiency than that of unsorted cells and 2.3-fold higher than cells sorted based on a single genetic marker, NrlGFP, expression. These proof-of-principle studies show that transplantation competent photoreceptor precursor cells can be efficiently isolated from a heterogeneous mix of cells using cell surface antigens without loss of viability for the purpose of retinal stem cell therapy. Refinement of the selection of donorphotoreceptor precursor cells can increase the number of integrated photoreceptor cells,which is a prerequisite for the restoration of sight.

Metabolic syndrome and coronary heart disease among Spanish male workers: a case-control study of MESYAS.

BACKGROUND AND AIMS: In Spain, the incidence of coronary heart disease is below that expected based on the burden of classic cardiovascular risk factors present in the population. Whether the risk associated with metabolic syndrome is lower in Spain deserves to be investigated. This study evaluates the association of incident clinical coronary heart disease with metabolic syndrome and each of its individual defining components in a sample of Spanish working males. METHODS AND RESULTS: Among the workers of a factory (MESYAS registry), 208 incident cases of coronary heart disease (between 1981 and 2005) were age-matched with 2080 healthy workers visited in 2004-2005. Metabolic syndrome was characterized using modified criteria of the joint consensus definition (2009). Metabolic syndrome was strongly associated with coronary heart disease (OR = 4.03; 95% CI: 2.98, 5.45) and the risk seemed to be fully explained by metabolic syndrome components (OR = 0.84, p = 0.54 after adjustment). Odds ratios for the independent effects of the diagnostic criteria were: hypertriglyceridemia (OR = 3.39, p < 0.001), hyperglycemia (OR = 2.70, p < 0.001), low HDL cholesterol (OR = 2.35, p < 0.001), hypertension (OR = 1.49, p = 0.016) and overweight (OR = 1.07, p = 0.678). Young workers showed a higher risk associated with metabolic syndrome. CONCLUSION: The risk associated with metabolic syndrome is fully explained by its components considered independently. The risk of coronary heart disease in a Spanish male working population is considerably increased among those with metabolic syndrome, by a factor similar to that described for other countries. Public health measures to prevent a rise in the prevalence of metabolic syndrome are advisable to minimize cardiovascular disease rate in Spain.

Circadian parameter as a possible indicator of gait performance and daily activity levels in chronic stroke survivors.

Stroke recovery is a critical public health problem. It involves cognitive and physical impairments leading to physical inactivity. In addition, there are also sleep problems and disruption of the circadian rhythm. In this paper, we analyze the relation between cognitive impairments level, walking performance, sleep and circadian parameters of a group of chronic stroke survivors. Five stroke survivors enrolled in the study. The clinical assessments were: cognitive function (MoCA), the 10 meter-walking test (10MWT), and the 6 minutes walking test (6MWT). The circadian rhythm parameters were obtained for at least one week wearing the actimeters. There was a positive correlation between the duration of the main sleep episode and the MoCA scores. There were significant correlations between the 10MWT and 6MWT and the circadian parameters that were positive for most active period (M10), Interdaily stability (IS), Mesor and Amplitude and negative for intradaily variability (IV). Our results indicate that gait speed and resistance are correlated to circadian rhythm synchronization, lower rhythm variability and more defined activity-rest episodes. This preliminary study underscores the importance of including a continuous measure of sleep and activity cycles in the assessment of stroke survivors.

Elbow movement estimation based on EMG with NARX Neural Networks.

The use of the electrical activity from the muscles may provide a natural way to control exoskeletons or other robotic devices seamlessly. The major challenges to achieve this goal are human motor redundancy and surface electromyography (sEMG) variability. The goal of this work is to find a feature extraction and classification procedures to estimate accurately elbow angular trajectory by means of a NARX Neural Network. The processing time-step should be small enough to make it feasible its further use for online control of an exoskeleton. In order to do so we analysed the Biceps and Triceps Brachii data from an elbow flexo-extension Coincident Timing task performed in the horizontal plane. The sEMG data was pre-processed and its energy was divided in five frequency intervals that were fed to a Nonlinear Auto Regressive with Exogenous inputs (NARX) Neural Network. The estimated angular trajectory was compared with the measured one showing a high correlation between them and a RMSE error maximum of 7 degrees. The procedure presented here shows a reasonably good estimation that, after training, allows real-time implementation. In addition, the results are encouraging to include more complex tasks including the shoulder joint.

EMG-Based Control in a Test Platform for Exoskeleton with One Degree of Freedom.

There are several efforts to use the electrical signals generated by the human muscles to control virtual or even physical devices. It is expected that, the development of this method will provide a natural way to control these devices, requiring little user training, depending on the task complexity. With respect to the control of exoskeletons from the electric signals generated by the muscles, it is desirable that the exoskeleton acts in synergy with the user using surface electromyography (sEMG) signals to detect user intentions. One of the challenges of this approach is the variability of the sEMG signals due to factors such as electrode positioning and conditions of the volunteer at the time of acquisition. In previous work, a procedure based on an Autoregressive with Exogenous Input (ARX) linear model was developed to translate sEMG from biceps, triceps and brachioradialis muscles to elbow joint angle. In this work, we developed a method based on a Genetic Algorithm (GA) to update the ARX model coefficients online to minimize the training periods and we have used the EMG signals to control a one-degree of freedom exoskeleton.vThe GA was able to obtain ARX model coefficients that generate the joint angle reference from the EMG signals. In addition, the joint angle references generated from the offline sEMG from three muscles via an ARX model were used to control a device. At this point we are carrying out tests with the exoskeleton using real-time signals from sEMG.

Behavioural characterisation of chronic unpredictable stress based on ethologically relevant paradigms in rats.

The chronic unpredictable stress (CUS) paradigm is extensively used in preclinical research. However, CUS exhibits translational inconsistencies, some of them resulting from the use of adult rodents, despite the evidence that vulnerability for many psychiatric disorders accumulates during early life. Here, we assessed the validity of the CUS model by including ethologically-relevant paradigms in juvenile rats. Thus, socially-isolated (SI) rats were submitted to CUS and compared with SI (experiment 1) and group-housed controls (experiment 1 and 2). We found that lower body-weight gain and hyperlocomotion, instead of sucrose consumption and preference, were the best parameters to monitor the progression of CUS, which also affected gene expression and neurotransmitter contents associated with that CUS-related phenotype. The behavioural characterisation after CUS placed locomotion and exploratory activity as the best stress predictors. By employing the exploratory factor analysis, we reduced each behavioural paradigm to few latent variables which clustered into two general domains that strongly predicted the CUS condition: (1) hyper-responsivity to novelty and mild threats, and (2) anxiety/depressive-like response. Altogether, the analyses of observable and latent variables indicate that early-life stress impairs the arousal-inhibition system leading to augmented and persistent responses towards novel, rewarding, and mildly-threatening stimuli, accompanied by lower body-weight gain.

Assessment of postural control after sleep deprivation with a low-cost portable force plate.

It is well-known that acute sleep deprivation affects negatively postural control. The analysis of sleep quality during long periods and its impact on motor control and learning performance are crucial aspects of human health. Nevertheless, there is conflicting evidence regarding which postural control variables are more prone to change due to sleep deprivation. Moreover, very few clinicians have at their disposal expensive force plates to measure such variables, so the use of a low-cost portable device could be very interesting. Therefore, we aimed to identify which posture control variables, obtained from a low-cost plate, are more sensitive to sleep deprivation. In order to do so, we have performed a set of experiments with volunteers before and after a night without sleep. Eight participants took part of the study and had their balance measured by a Wii Balance Board before and after one night of sleep deprivation. They were asked to keep a quiet stance on top of the plate with their eyes open and closed, in a balanced design. The main results showed that, regardless the visual information, sleep deprivation has deepest impact on the anterior-posterior center of pressure displacement. Sleep deprivation without visual information had a more pronounced (large effect size) impact on the mean sway in the anterior-posterior direction and its distribution variation. The information that sleep deprivation has a more meaningful impact on anterior-posterior center of pressure excursion may help clinicians and healthcare professionals to better deal with its implications.

Study of the motion artefacts of skin-mounted inertial sensors under different attachment conditions.

A common problem shared by accelerometers, inertial sensors and any motion measurement method based on skin-mounted sensors is the movement of the soft tissues covering the bones. The aim of this work is to propose a method for the validation of the attachment of skin-mounted sensors. A second-order (mass-spring-damper) model was proposed to characterize the behaviour of the soft tissue between the bone and the sensor. Three sets of experiments were performed. In the first one, different procedures to excite the system were evaluated to select an adequate excitation stimulus. In the second one, the selected stimulus was applied under varying attachment conditions while the third experiment was used to test the model. The heel drop was chosen as the excitation method because it showed lower variability and could discriminate between different attachment conditions. There was, in agreement with the model, a trend to increase the natural frequency of the system with decreasing accelerometer mass. An important result is the development of a standard procedure to test the bandwidth of skin-mounted inertial sensors, such as accelerometers mounted on the skin or markers heavier than a few grams.

Cochlear obliteration following a translabyrinthine approach and its implications in cochlear implantation.

The most frequent sequelae following a translabyrinthine approach for vestibular schwannoma resection is complete hearing loss on the affected side. Such patients could benefit from a cochlear implant, provided that two essential requisites are met before surgery: a preserved cochlear nerve and a patent cochlea to accommodate the electrode array. The goal of our study is to determine the prevalence and extent of cochlear ossification following a translabyrinthine approach. Postoperative MRI of 41 patients were retrospectively reviewed. Patients were classified according to the degree of cochlear obliteration into three groups (patent cochlea, partially obliterated cochlea and totally obliterated cochlea). The interval between surgery and the first MRI was studied as well as its relationship with the rate of cochlear ossification. At first postoperative MRI (mean interval of 20 months), 78% of patients showed some degree of cochlear ossification. Differences were found in the time interval between surgery and first MRI for each group, showing a smaller interval of time the patent cochlea group (p > 0.05). When MRI was performed before the first year after surgery, a larger rate of patent cochlea was found (p > 0.05). The present study suggests that cochlear ossification is a time-depending process, whose grounds are still to be defined.

Daytime sleepiness affects gait auditory synchronization ability.

Sleep disturbances in modern life lead to cognitive and motor performance impairments in everyday tasks such as gait. The most common symptom of these disturbances is daytime sleepiness, which can be assessed by questionnaires such as the Epworth Sleep Scale (ESS). The ESS evaluates sleep health and daytime dysfunction. The goal of this study is to assess the influence of sleepiness on a motorauditory synchrony task, rhythmed gait. High and low sleepiness clusters were formed based on the participants ESS scores. Walking on a treadmill, two different rhythmic auditory stimulus conditions were set with a metronome: isochronous and non-isochronous. Reflective markers on both heels with seven infrared cameras were used to assess the difference between footfall and metronome beep, what is named synchronization error (SE). There was a tendency to anticipate the beep in the HS group when compared to the LS group only in the non-isochronous stimulus condition that was statistically significant. Sleep disturbances that generate daytime sleepiness may bring detrimental effects on brain areas that could be responsible for the real-time adjustment of gait and sustained attention. These impairments may be responsible for the larger synchronization error with larger relative phase of the group with high sleepiness. More studies are necessary involving other parameters of sleep and gait to identify sleep disturbances through gait analysis.

Elbow Joint Angle Estimation with Surface Electromyography Using Autoregressive Models.

This paper presents a method to estimate the elbow joint angle from surface electromyography (sEMG) measurements of biceps, triceps and brachioradialis. This estimation is of major importance for the design of human robot interfaces based on sEMG. It is also relevant to model the muscular system and to design biomimetic mechanisms. However, the processing and interpretation of electromyographic signals is challenging due to nonlinearities, unmodeled muscle dynamics, noise and interferences. In order to determine an estimation model and a calibration procedure for the model parameters, a set of experiments were carried out with six subjects. The experiments consisted of series of continuous (cyclical) and discrete elbow flexo-extensions with three different loads (i.e. 0 kg, 1.5kg and 3 kg). The sEMG data from the biceps brachii, triceps brachii and brachioradialis and the joint angle were recorded. Four different modeling techniques were evaluated: State Space (SS), Autoregressive with Exogenous Input (ARX), Autoregressive Moving-Average with Exogenous Input (ARMAX), Autoregressive Integrated Moving-Average with Exogenous Input (ARIMAX). After the model was selected, a second experiment was performed in order to validate the estimation procedure. The results show a procedure to estimate the EMG-to-angle relation with high correlation and low meansquare- root errors with respect to the measured angle data.

Two-dose daclizumab, tacrolimus, mycophenolate mofetil, and steroid-free regimen in de novo cardiac transplant recipients: early experience.

BACKGROUND: Tacrolimus (TAC) with mycophenolate mofetil (MMF) and a steroid-free regimen seems to have good efficacy in preventing acute rejection in cardiac transplant recipients, although concern exists about nephrotoxicity. Induction therapy with Daclizumab seems to give protection without side effects. Data are lacking about the outcome of 2-dose Daclizumab+TAC+MMF and a steroid-free regimen. MATERIALS AND METHODS: We retrospectively reviewed 28 consecutive de novo heart transplantations performed at a single center between January 2001 and June 2006. Patients received induction therapy with 2-dose Daclizumab. Maintenance immunosuppression included TAC, MMF, and prednisone during the first 6 months. The endpoints were the incidence of acute rejection, patient and graft survival, and clinical tolerability. RESULTS: Among 28 patients of mean age 57 +/- 9 years, 2 subjects (7%) died in the perioperative period due to infections. The mean follow-up was 2.8 +/- 1.5 years. There were no late deaths. Six patients experienced acute rejection (International Society of Heart and Lung Transplantation [ISHLT] >or=3A) that required treatment during the first 3 months. At follow-up, only 3 patients (>or=3A) required treatment. Mean creatinine level increased from 1.08 +/- 0.37 at baseline to 1.08 +/- 0.41 at 1 year (n = 23; P = not significant [NS]) to 1.39 +/- 0.68 (n = 13; P < .05) at 4 years, 1.65 +/- 0.51 (n = 8; P < .05) at 5 years. No patient required replacement therapy. CONCLUSIONS: A steroid-free protocol with 2-dose Daclizumab induction therapy and maintenance with TAC and MMF seemed to be safe to prevent acute rejection. Creatinine levels were slightly but significantly increased.

Impact on renal function of the use of sirolimus in cardiac transplantation.

INTRODUCTION: During the last few years sirolimus has been introduced as an alternative to preserve renal function in transplant recipients receiving calcineurin inhibitors. MATERIALS AND METHODS: We reviewed our results on the use of sirolimus in cardiac transplant recipients. RESULTS: Twenty-seven patients with an average age of 63 years received sirolimus. The average time after transplantation was 73.4 +/- 58.9 months and the average follow-up was 31.7 +/- 18.01 months. Sirolimus was prescribed in 37% of cases due to chronic renal failure (CRF), 14.8% because of cardiac allograft vasculopathy (CAV), 11.1% for tumors, 22.2% de novo, 7.4% for CRF and tumor, and 7.4% for CRF and CAV. Among the patients with CRF (n = 14), there were 5 (35%) on dialysis at the moment of starting the treatment and 1 was retired from dialysis. The other 4 (28.5%) patients had to be treated with dialysis after starting the treatment. In all, 42.8% of the patients with nephropathy maintained stable renal function or improved. Among the 17 (63%) patients who did not require dialysis, there was no significant change in renal function after 6 months or 1, 2, and 3 years follow-up. CONCLUSIONS: The use of sirolimus in cardiac transplantation maintains stable renal function in the majority of patients in the medium term.

Posture control and complex arm coordination: analysis of multijoint coordinative movements and stability of stance.

The authors addressed the interactions between control of bimanual multijoint coordination tasks and posture. Participants (N = 6) performed 8 coordination patterns that differed in degree of complexity by using their bilateral elbows and wrists under 3 scaled motion speeds while standing on 2 force plates. Results indicated that producing complex bimanual multijoint coordinative tasks affected postural sway, thus resulting in an increase of sway activity. Behavioral as well as mechanical factors accounted for the increased disturbance in postural sway. Those findings suggest that performing complex coordination tasks disrupts postural control in normal young adults.

Biomechanical procedure to assess sleep restriction on motor control and learning.

The analysis of sleep quality during long periods and its impact on motor control and learning performance are crucial aspects for human health. The aim of this study is to analyze effects of chronic sleep restriction on motor performance. It is intended to establish motor control indicators in sleep quality analysis. A wearable actigraphy that records accelerometry, ambient light, and body temperature was used to monitor the sleep habits of 12 healthy subjects for two weeks before performing motor control and learning tests. The day of the motor test, the subjects filled two questionnaires about the quality of sleep (Pittsburgh Sleep Quality Index - PSQI) and sleepiness (Epworth Sleepiness Scale - ESS). Afterwards they performed a coincident timing task that consisted of hitting a virtual target falling on the screen with the hand. An elbow flexion in the horizontal plane had to be performed on the correct time to reach the real target on a table at the same time as the virtual target on the screen. The subjects performed three sets of acquisition and transfer blocks of the coincident timing task. The subjects were clustered in two groups based on the PSQI and ESS scores. Actigraphy and motor control parameters (L5, correct responses, time variance) were compared between groups and experimental sets. The group with better sleep parameters did show a constant performance across blocks of task acquisition while the bad sleeper group improved from the first to the second acquisition block. Despite of this improvement, their performance is not better than the one of the good sleepers group. Although the number of subjects is low and it should be increased, these results indicate that the subjects with better sleep converged rapidly to a high level of performance, while the worse sleepers needed more trials to learn the task and their performance was not superior to the other group.

First case of Nocardia amamiensis pulmonary infection in Mexico.

We report a case of Nocardia amamiensis pulmonary infection in a 43-year-old immunocompromised woman. The patient was treated with imipenem/cilastatin and trimethoprim/sulfamethoxazole and had a favourable outcome. It is important that laboratories perform species identification to understand the epidemiology and susceptibility patterns of the different Nocardia spp.