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Estudou-se a marcação imunohistoquímica para o MIB-1 (Ki-67) em 24 carcinomas da córtex adrenal. A idade dos pacientes variou de 1 a 53 anos, com mediana de 7,5. Excetuando 1 paciente com doença avançada, todos foram submetidos à adrenalectomia. O tempo de seguimento variou de 1 a 13 anos. No momento do diagnóstico 7/24 pacientes já apresentavam metástases e no decorrer do seguimento 3 pacientes adicionais as desenvolveram. A média do peso tumoral dos sobreviventes livres da doença foi de 400± 687 g enquanto que nos falecidos ou com neoplasia após a cirurgia foi de 508± 335 g, diferença essa sem significado estatístico. Tumores maiores que 200 g mostraram pior prognóstico do que aqueles com peso menor (p = 0,01). Os valores médios do índice mitótico (IM) para tumores em estádio I, II, III e IV foram respectivamente: 17,4± 10,0, 12,4± 11,2, 18,3± 14,0 e 28,3± 21,0. A comparação das médias revelou p>0,2. Os valores médios do índice de proliferação(IP) determinado pelo MIB-1 nos estádios I, II, III e IV foram respectivamente: 201,4± 131,0, 179,6± 98,7, 129,1± 170,2 e 124,0± 99,4. A comparação estatística dessas médias mostrou p>0,3. Valores do IP maiores que 20 estão associados a pior prognóstico. O IP não mostrou relação com o prognóstico.

Chronic Obstructive Pulmonary Disease (COPD) is a pathological condition of the respiratory system characterized by chronic airflow obstruction, associated with changes in the lung parenchyma (pulmonary emphysema), bronchi (chronic bronchitis) and bronchioles (small airways disease). In the last years, the importance of phenotyping and endotyping COPD patients has strongly emerged. Metabolomics refers to the study of metabolites (both intermediate or final products) and their biological processes in biomatrices. The application of metabolomics to respiratory diseases and, particularly, to COPD started more than one decade ago and since then the number of scientific publications on the topic has constantly grown. In respiratory diseases, metabolomic studies have focused on the detection of metabolites derived from biomatrices such as exhaled breath condensate, bronchoalveolar lavage, and also plasma, serum and urine. Mass Spectrometry and Nuclear Magnetic Resonance Spectroscopy are powerful tools in the precise identification of potentially prognostic and treatment response biomarkers. The aim of this article was to comprehensively review the relevant literature regarding the applications of metabolomics in COPD, clarifying the potential clinical utility of the metabolomic profile from several biologic matrices in detecting biomarkers of disease and prognosis for COPD. Meanwhile, a complete description of the technological instruments and techniques currently adopted in the metabolomics research will be described., Competing Interests: The authors declare no conflicts of interest.

Niemann-Pick Disease (NPD) is a rare autosomal recessive disease belonging to lysosomal storage disorders. Three types of NPD have been described: NPD type A, B, and C. NPD type A and B are caused by mutations in the gene SMPD1 coding for sphingomyelin phosphodiesterase 1, with a consequent lack of acid sphingomyelinase activity. These diseases have been thus classified as acid sphingomyelinase deficiencies (ASMDs). NPD type C is a neurologic disorder due to mutations in the genes NPC1 or NPC2 , causing a defect of cholesterol trafficking and esterification. Although all three types of NPD can manifest with pulmonary involvement, lung disease occurs more frequently in NPD type B, typically with interstitial lung disease, recurrent pulmonary infections, and respiratory failure. In this sense, bronchoscopy with broncho-alveolar lavage or biopsy together with high-resolution computed tomography are fundamental diagnostic tools. Although several efforts have been made to find an effective therapy for NPD, to date, only limited therapeutic options are available. Enzyme replacement therapy with Olipudase α is the first and only approved disease-modifying therapy for patients with ASMD. A lung transplant and hematopoietic stem cell transplantation are also described for ASMD in the literature. The only approved disease-modifying therapy in NPD type C is miglustat, a substrate-reduction treatment. The aim of this review was to delineate a state of the art on the genetic basis and lung involvement in NPD, focusing on clinical manifestations, radiologic and histopathologic characteristics of the disease, and available therapeutic options, with a gaze on future therapeutic strategies.

Purpose: To produce and validate an Italian version of the Lymphedema Life Impact Scale version 2 (LLISv2-It), a tool measuring the impact of lymphedema on health-related quality of life, and investigate its main psychometric characteristics., Methods: After translation and cross-cultural adaptation of the LLISv2, we administered it to 156 subjects with secondary lymphedema (upper or lower limb), together with (depending on the limb involved) either the Disabilities of the Arm, Shoulder and Hand questionnaire (DASH) or the Lower Extremity Functional Scale (LEFS). We analyzed the performance of LLISv2-It using Classical Test Theory and Rasch methods., Results: Cronbach alpha was 0.89. Item fit statistics indicated that item #12 was underfitting (Infit MnSQ = 1.56; Outfit MnSq = 1.75). Test-retest reliability of the 17-item version (without item #18, not scored, according to the original authors) and of a 16-item version (without #12 and #18-a unidimensional item set) were both excellent (ICC2.1 = 0.93; 95%). The minimum detectable change was 8.9 points. The correlation of LLISv2-It with DASH and LEFS was r  = 0.81 and -0.57, respectively., Conclusions: The Italian version of the LLISv2 is reliable and valid. To make the tool more psychometrically sound, we recommend to not calculate items #12 and #18 in the total score., IMPLICATIONS FOR REHABILITATIONThe Lymphedema Life Impact Scale (LLISv2) is particularly useful as a validated condition-specific health-related quality of life measure for patients with chronic edema/lymphedema. We have validated its Italian version (LLISv2-It).We also examined the psychometric properties of the LLISv2 in individuals with secondary lymphedema of either upper or lower limb, using both classical test theory and Rasch techniques, to provide insights for further refinement of the questionnaire.Based on our results, we recommend not to count item #12 in the total score (besides #18, as already suggested by the original authors). Moreover, we determined that the minimum detectable change of the LLISv2 was 9 points. This information is helpful in clinical practice to understand if the change in score is not due to chance (measurement error).Although further research is warranted, the present validation study of the LLISv2 increases the confidence in the metric quality and clinical utility of the scale to assess lymphedema-specific health-related quality of life in individuals with secondary lymphedema of either upper or lower limb.

Background: Walking impairments are a common consequence of neurological disorders and are assessed with clinical scores that suffer from several limitations. Robot-assisted locomotor training is becoming an established clinical practice. Besides training, these devices could be used for assessing walking ability in a controlled environment. Here, we propose an adaptive assist-as-needed (AAN) control for a treadmill-based robotic exoskeleton, the Lokomat, that reduces the support of the device (body weight support and impedance of the robotic joints) based on the ability of the patient to follow a gait pattern displayed on screen. We hypothesize that the converged values of robotic support provide valid and reliable information about individuals' walking ability., Methods: Fifteen participants with spinal cord injury and twelve controls used the AAN software in the Lokomat twice within a week and were assessed using clinical scores (10MWT, TUG). We used a regression method to identify the robotic measure that could provide the most relevant information about walking ability and determined the test-retest reliability. We also checked whether this result could be extrapolated to non-ambulatory and to unimpaired subjects., Results: The AAN controller could be used in patients with different injury severity levels. A linear model based on one variable (robotic knee stiffness at terminal swing) could explain 74% of the variance in the 10MWT and 61% in the TUG in ambulatory patients and showed good relative reliability but poor absolute reliability. Adding the variable 'maximum hip flexor torque' to the model increased the explained variance above 85%. This did not extend to non-ambulatory nor to able-bodied individuals, where variables related to stance phase and to push-off phase seem more relevant., Conclusions: The novel AAN software for the Lokomat can be used to quantify the support required by a patient while performing robotic gait training. The adaptive software might enable more challenging training conditions tuned to the ability of the individuals. While the current implementation is not ready for assessment in clinical practice, we could demonstrate that this approach is safe, and it could be integrated as assist-as-needed training, rather than as assessment., Trial Registration: ClinicalTrials.gov Identifier: NCT02425332., (© 2023. BioMed Central Ltd., part of Springer Nature.)

Most robotic gait assisted devices are designed to provide constant assistance during the training without taking into account each patient's functional ability. The Lokomat offers an assist-as-needed control via the integrated exercise "Adaptive Gait Support" (AGS), which adapts the robotic support based on the patient's abilities. The aims of this study were to examine the feasibility and characteristics of the AGS during long-term application. Ten patients suffering from neurological diseases underwent an 8-week Lokomat training with the AGS. They additionally performed conventional walking tests and a robotic force measurement. The difference between robotic support during adaptive and conventional training and the relationship between the robotic assessment and the conventional walking and force tests were examined. The results show that AGS is feasible during long-term application in a heterogeneous population. The support during AGS training in most of the gait phases was significantly lower than during conventional Lokomat training. A relationship between the robotic support level determined by the AGS and conventional walking tests was revealed. Moreover, combining the isometric force data and AGS data could divide patients into clusters, based on their ability to generate high forces and their level of motor control. AGS shows a high potential in assessing patients' walking ability, as well as in providing challenging training, e.g., by automatically adjusting the robotic support throughout the whole gait cycle and enabling training at lower robotic support.

Purpose: There is growing interest in measures that assess upper-limb lymphedema after breast cancer. Since no validated Italian version of the Lymphedema Quality of Life Questionnaire for upper limbs (LYMQOL-UL) exists, we aimed to culturally adapt and validate an Italian version (LYMQOL-UL-IT) in order to allow its use in Italian patients., Materials and Methods: The LYMQOL-UL-IT was developed by means of forward-backward translation, review by an expert committee and a test of the pre-final version to evaluate its comprehensibility. The psychometric testing included reliability by internal consistency (Cronbach's alpha) and test-retest reliability (intraclass correlation coefficient, ICC (2.1) ), measurement error by calculating the minimum detectable change (MDC 95 ), construct validity by confirmatory factor analysis, and evaluation of a priori hypotheses about the correlations between the four LYMQOL-UL domains, single items of the LYMQOL-UL, and measures of health-related quality of life and pain intensity (Spearman's rank correlation coefficient)., Results: The consensus-based version of LYMQOL-UL-IT was administered to 139 patients with upper-limb stable secondary iatrogenic lymphedema after breast cancer. The adapted questionnaire was well accepted as it was completed in less than 10 min, without missing answers or comprehension problems. Internal consistency was acceptable (α = 0.92-0.95). Test-retest reliability was good-to-excellent (ICC (2.1)  = 0.73-0.96). The MDC 95 for the four domains of the questionnaire was as follows: 0.64 scale points for Function, 0.40 for Appearance, 0.53 for Symptoms, and 0.81 for Mood. Factor analysis confirmed a 4-dimensional structure as originally conceived and the a priori hypotheses were met., Conclusion: The LYMQOL-UL-IT is reliable, sensitive to change and valid in patients with upper-limb stable secondary iatrogenic lymphedema after breast cancer. It can be used for clinical and research purposes.Implications for rehabilitationLymphedema is a frequently unnoticed clinical condition that not only impacts physical functioning but often restricts the health-related quality of life in breast cancer survivors.The Italian version of the Lymphedema Quality of Life Questionnaire for upper limbs (LYMQOL-UL-IT) is reliable, sensitive to change and valid in patients with upper-limb lymphedema after breast cancer.The LYMQOL-UL-IT tool can be recommended for clinical and research purposes.

Robotic algorithms that augment movement errors have been proposed as promising training strategies to enhance motor learning and neurorehabilitation. However, most research effort has focused on rehabilitation of upper limbs, probably because large movement errors are especially dangerous during gait training, as they might result in stumbling and falling. Furthermore, systematic large movement errors might limit the participants' motivation during training. In this study, we investigated the effect of training with novel error modulating strategies, which guarantee a safe training environment, on motivation and learning of a modified asymmetric gait pattern. Thirty healthy young participants walked in the exoskeletal robotic system Lokomat while performing a foot target-tracking task, which required an increased hip and knee flexion in the dominant leg. Learning the asymmetric gait pattern with three different strategies was evaluated: (i) No disturbance: no robot disturbance/guidance was applied, (ii) haptic error amplification: unsafe and discouraging large errors were limited with haptic guidance, while haptic error amplification enhanced awareness of small errors relevant for learning, and (iii) visual error amplification: visually observed errors were amplified in a virtual reality environment. We also evaluated whether increasing the movement variability during training by adding randomly varying haptic disturbances on top of the other training strategies further enhances learning. We analyzed participants' motor performance and self-reported intrinsic motivation before, during and after training. We found that training with the novel haptic error amplification strategy did not hamper motor adaptation and enhanced transfer of the practiced asymmetric gait pattern to free walking. Training with visual error amplification, on the other hand, increased errors during training and hampered motor learning. Participants who trained with visual error amplification also reported a reduced perceived competence. Adding haptic disturbance increased the movement variability during training, but did not have a significant effect on motor adaptation, probably because training with haptic disturbance on top of visual and haptic error amplification decreased the participants' feelings of competence. The proposed novel haptic error modulating controller that amplifies small task-relevant errors while limiting large errors outperformed visual error augmentation and might provide a promising framework to improve robotic gait training outcomes in neurological patients.

Assist-as-needed (AAN) algorithms for the control of lower extremity rehabilitation robots can promote active participation of patients during training while adapting to their individual performances and impairments. The implementation of such controllers requires the adaptation of a control parameter (often the robot impedance) based on a performance (or error) metric. The choice of how an adaptive impedance controller is formulated implies different challenges and possibilities for controlling the patient's leg movement. In this paper, we analyze the characteristics and limitations of controllers defined in two commonly used formulations: joint and end-point space, exploring especially the implementation of an AAN algorithm. We propose then, as a proof-of-concept, an AAN impedance controller that combines the strengths of working in both spaces: a hybrid joint/end-point impedance controller. This approach gives the possibility to adapt the end-point stiffness in magnitude and direction in order to provide a support that targets the kinematic deviations of the end-point with the appropriate force vector. This controller was implemented on a two-link rehabilitation robot for gait training-the Lokomat®Pro V5 (Hocoma AG, Switzerland) and tested on 5 able-bodied subjects and 1 subject with Spinal Cord Injury. Our experiments show that the hybrid controller is a feasible approach for exoskeleton devices and that it could exploit the benefits of the end-point controller in shaping a desired end-point stiffness and those of the joint controller to promote the correct angular changes in the trajectories of the joints. The adaptation algorithm is able to adapt the end-point stiffness based on the subject's performance in different gait phases, i.e., the robot can render a higher stiffness selectively in the direction and gait phases where the subjects perform with larger kinematic errors. The proposed approach can potentially be generalized to other robotic applications for rehabilitation or assistive purposes., (Copyright © 2018 Maggioni, Reinert, Lünenburger and Melendez-Calderon.)

Many robotic devices that are used for therapy or as assistive devices rely on pre-defined reference trajectories to assist the user's movements. Fixed pre-defined trajectories force the user to adapt to unnatural movement patterns which may be detrimental to rehabilitation outcomes. We propose a database-driven approach to adapt the reference trajectory of robotic training devices that rely on cyclic motion such as walking. Dynamic time warping is used to compare the measured pattern with a database of pre-approved safe trajectories; the best matching pattern is selected from the database and used for the next movement sequence. The proposed approach was evaluated in computer simulations and a bioinspired robotic test bench. Our proposed method reduced the RMS error between individual user trajectories and the supplied reference, even in the presence of measurement noise.

Heterogeneous results have been obtained in the relationship between serum uric acid (SUA) and target organ damage (TOD) in patients with hypertension. Clinic blood pressure, SUA, and cardiac, arterial (carotid and aortic), and renal TOD were assessed in 762 consecutive patients with hypertension. Hyperuricemia was defined as an SUA >7.0 in men and >6.0 mg/dL in women. Men with hyperuricemia compared with those with normal SUA showed lower estimated glomerular filtration rates and E/A ratios and a higher prevalence of carotid plaques. Women with hyperuricemia showed lower estimated glomerular filtration rates and E/A ratios and a higher intima-media thickness. Except for pulse wave velocity, all TODs significantly correlated with SUA. However, at multivariate analysis, only estimated glomerular filtration rate was significantly determined by SUA. Our data provide evidence on the role of SUA in the development of TOD only in the case of renal alteration. It is likely that SUA may indirectly act on the other TODs through the increase in blood pressure and the decrease in glomerular filtration rate., (©2017 Wiley Periodicals, Inc.)

Clinically useful and efficient assessment of balance during standing and walking is especially challenging in patients with neurological disorders. However, rehabilitation robots could facilitate assessment procedures and improve their clinical value. We present a short overview of balance assessment in clinical practice and in posturography. Based on this overview, we evaluate the potential use of robotic tools for such assessment. The novelty and assumed main benefits of using robots for assessment are their ability to assess 'severely affected' patients by providing assistance-as-needed, as well as to provide consistent perturbations during standing and walking while measuring the patient's reactions. We provide a classification of robotic devices on three aspects relevant to their potential application for balance assessment: 1) how the device interacts with the body, 2) in what sense the device is mobile, and 3) on what surface the person stands or walks when using the device. As examples, nine types of robotic devices are described, classified and evaluated for their suitability for balance assessment. Two example cases of robotic assessments based on perturbations during walking are presented. We conclude that robotic devices are promising and can become useful and relevant tools for assessment of balance in patients with neurological disorders, both in research and in clinical use. Robotic assessment holds the promise to provide increasingly detailed assessment that allows to individually tailor rehabilitation training, which may eventually improve training effectiveness.

The assessment of sensorimotor functions is extremely important to understand the health status of a patient and its change over time. Assessments are necessary to plan and adjust the therapy in order to maximize the chances of individual recovery. Nowadays, however, assessments are seldom used in clinical practice due to administrative constraints or to inadequate validity, reliability and responsiveness. In clinical trials, more sensitive and reliable measurement scales could unmask changes in physiological variables that would not be visible with existing clinical scores.In the last decades robotic devices have become available for neurorehabilitation training in clinical centers. Besides training, robotic devices can overcome some of the limitations in traditional clinical assessments by providing more objective, sensitive, reliable and time-efficient measurements. However, it is necessary to understand the clinical needs to be able to develop novel robot-aided assessment methods that can be integrated in clinical practice.This paper aims at providing researchers and developers in the field of robotic neurorehabilitation with a comprehensive review of assessment methods for the lower extremities. Among the ICF domains, we included those related to lower extremities sensorimotor functions and walking; for each chapter we present and discuss existing assessments used in routine clinical practice and contrast those to state-of-the-art instrumented and robot-aided technologies. Based on the shortcomings of current assessments, on the identified clinical needs and on the opportunities offered by robotic devices, we propose future directions for research in rehabilitation robotics. The review and recommendations provided in this paper aim to guide the design of the next generation of robot-aided functional assessments, their validation and their translation to clinical practice.

Flaxseeds are both a food ingredient and a natural source of antioxidants (e.g. lignans, PUFAs) and pro-oxidant contaminants (e.g. cadmium): the variable mixture of anti- and pro-oxidant substances may impact on the redox homeostasis of flaxseed-enriched foods. The antioxidant power is studied here as biochemical activity of flaxseeds in white wheat bread and as endpoint for possible screening of anomalous variations of bioactive mixtures (antioxidants vs. prooxidants) in food matrices. A bioprobe assay based on the superoxide dismutase (SOD) enzyme (6 channels of the multiprobe bioelectronic platform BEST) was performed on white wheat bread with and without flaxseeds. Nine BEST channels were simultaneously used for validation and monitoring of measuring conditions (temperature, pH, conductivity). Findings were compared with quantitative analysis of antioxidants and pro-oxidant contaminants. Organic and aqueous extracts of both bread types were examined in parallel. The SOD-probe detected the difference in antioxidant power given by 10% flaxseed, thus supporting the use of antioxidant power detected by bioenzymatic screening as sensitive biochemical endpoint. Mixtures of bioactive molecules in foods generate biochemical activities that can be monitored as time-effective indicators of invariability, which is pivotal in the daily control of anomalies in food production and therefore in the protection of consumers' health., (Copyright © 2016 Elsevier Ltd. All rights reserved.)

Aims/hypothesis: We investigated the significance of microangiopathy in the development of foot ulcer, which is still disputed., Methods: We assessed microangiopathy by histological analysis of the capillary ultrastructure using transmission electron microscopy and capillary density and arteriolar morphology in paraffin-embedded sections from the skin of type 2 diabetic patients: 30 neuroischaemic patients (Isc) revascularised with peripheral angioplasty and 30 neuropathic patients (Neu) with foot ulcer, compared with ten non-diabetic volunteers., Results: In the diabetic patients, capillaries in the dermal papillary layer were fewer (-22.2%, 159 ± 43 vs 205 ± 52 mm(2) in non-diabetic volunteers, p < 0.01). They also showed detrimental remodelling, with a 2.2-fold increase in capillary basement membrane thickness (3.44 ± 1.19 vs 1.53 ± 0.34 μm in non-diabetic volunteers, p < 0.001) and a 57.7% decrease in lumen area (14.6 ± 11.1 vs 34.7 ± 27.5 μm(2), p < 0.001). No differences were observed between the diabetic Isc or Neu patients. Isc were more prone to develop arteriolar occlusion than Neu (16.8 ± 6.9% vs 6.7 ± 3.7%, respectively, p < 0.001). No patient had been amputated at 30 days and healing time was significantly longer in Isc (180 ± 120 vs 64 ± 50 days in Neu, p < 0.001)., Conclusions/interpretation: Capillary microangiopathy is present in equal measure in neuroischaemic and neuropathic diabetic foot skin. The predominance of arteriolar occlusions with neuroischaemia indicated the existence of an additional 'small vessel disease' that did not affect an effective revascularisation and did not worsen the prognosis of major amputations but slowed the healing process of the neuroischaemic foot ulcer., Trial Registration: ClinicalTrials.gov NCT02610036.

Aims: We examined whether, in diabetic Ob/Ob mice, the dipeptidyl peptidase-4 (DPP-4) inhibitor (PKF275-055), an antihyperglycemic drug, that inhibits the biological inactivation of SDF-1 (stromal cell-derived factor-1), may increase endothelial progenitor cells (EPCs) mobilization and incorporation, which, in turn, may regenerate capillaries and reduce myocardial ischemia induced by strenuous exercise., Main Methods: Half of sixteen control and Ob/Ob mice and eight Ob/Ob mice treated with PKF275-055 for four weeks underwent a forced swim protocol. Oral glucose tolerance, circulating EPCs, capillary ultrastructure and density, hypoxic areas and SDF-1 localization in myocardium were measured., Key Findings: Ob/Ob mice were glucose intolerant, had a significant low number of circulating EPCs and myocardial capillaries compared to lean controls. The DPP-4 inhibitor significantly improved their glucose tolerance, doubled the number of circulating EPCs, stimulated the formation of functional vessels and SDF-1 localization in the endothelium of myocardial capillaries and arterioles. Cardiac hypoxia after forced swim in Ob/Ob mice was significantly reduced when they were treated with the DPP-4 inhibitor., Significance: DPP-4 inhibition may re-establish an adequate capillary network in the myocardium of diabetic Ob/Ob mice by the mobilization and SDF-1-mediated incorporation of EPCs and, consequently, reducing the susceptibility to myocardial ischemic injury provoked by strenuous exercise., (Copyright © 2016 Elsevier Inc. All rights reserved.)

Long-term repeated-dose toxicity is mainly assessed in animals despite poor concordance of animal data with human toxicity. Nowadays advanced human in vitro systems, eg, metabolically competent HepaRG cells, are used for toxicity screening. Extrapolation of in vitro toxicity to in vivo effects is possible by reverse dosimetry using pharmacokinetic modeling. We assessed long-term repeated-dose toxicity of bosentan and valproic acid (VPA) in HepaRG cells under serum-free conditions. Upon 28-day exposure, the EC50 values for bosentan and VPA decreased by 21- and 33-fold, respectively. Using EC(10) as lowest threshold of toxicity in vitro, we estimated the oral equivalent doses for both test compounds using a simplified pharmacokinetic model for the extrapolation of in vitro toxicity to in vivo effect. The model predicts that bosentan is safe at the considered dose under the assumed conditions upon 4 weeks exposure. For VPA, hepatotoxicity is predicted for 4% and 47% of the virtual population at the maximum recommended daily dose after 3 and 4 weeks of exposure, respectively. We also investigated the changes in the central carbon metabolism of HepaRG cells exposed to orally bioavailable concentrations of both drugs. These concentrations are below the 28-day EC(10) and induce significant changes especially in glucose metabolism and urea production. These metabolic changes may have a pronounced impact in susceptible patients such as those with compromised liver function and urea cycle deficiency leading to idiosyncratic toxicity. We show that the combination of modeling based on in vitro repeated-dose data and metabolic changes allows the prediction of human relevant in vivo toxicity with mechanistic insights., (© The Author 2015. Published by Oxford University Press on behalf of the Society of Toxicology. All rights reserved. For Permissions, please e-mail: journals.permissions@oup.com.)

Background: Even if movement abnormalities in dystonia are obvious on observation-based examinations, objective measures to characterize dystonia and to gain insights into its pathophysiology are still strongly needed. We hypothesize that motor abnormalities in childhood dystonia are partially due to the inability to suppress involuntary variable muscle activity irrelevant to the achievement of the desired motor task, resulting in the superposition of unwanted motion components on the desired movement. However, it is difficult to separate and quantify appropriate and inappropriate motor signals combined in the same muscle, especially during movement., Methods: We devise an innovative and practical method to objectively measure movement abnormalities during the performance of a continuous figure-eight writing task in 7 children with dystonia and 9 age-matched healthy controls. During the execution of a continuous writing task, muscle contractions should occur at frequencies that match the frequencies of the writing outcome. We compare the power spectra of kinematic trajectories and electromyographic signals of 8 upper limb muscles to separate muscle activity with the same frequency content of the figure-eight movement (task-correlated) from activity occurring at frequencies extraneous to the task (task-uncorrelated)., Results: Children with dystonia present a greater magnitude of task-uncorrelated muscle components. The motor performance achieved by children with dystonia is characterized by an overall lower quality, with high spatial and temporal variability and an altered trade-off between speed and accuracy., Conclusions: Findings are consistent with the hypothesis that, in childhood dystonia, the ability to appropriately suppress variable and uncorrelated elements of movement is impaired. Here we present a proof-of-concept of a promising tool to characterize the phenomenology of movement disorders and to inform the design of neurorehabilitation therapies.

Purpose: We have designed a desensitization program that gives good results and is cost effective for kidney-transplant patients who have a potential living donor, who are ABO incompatible (ABOi), and who may or may not have donor-specific alloantibodies (DSAs)., Methods: Desensitization at pretransplant is based on immunosuppressants (such as rituximab, tacrolimus, and mycophenolic acid) and apheresis to retrieve potentially detrimental isoagglutinins and DSAs from blood. In 2011, we implemented immunoadsorption (IA) instead of plasmapheresis in our center as part of the desensitization protocol. Because IA is very tedious and time-consuming, we decided to perform IA and hemodialysis (HD) in tandem instead of performing these methods sequentially. Herein, we report on more than 100 of these tandem procedures and compare the results to those obtained when IA was performed before a HD session., Results: The tandem process resulted in nursing time being cut by half: from almost 10h30 min/patient to 6 h/patient. When IA was performed alone, body-weight gain was 1 kg (range 0 to 1.75) and the median volume of plasma treated per session was 6500 mL (range 4500 to 10 600). Conversely, during a tandem IA + HD session, the net median weight gain was -1.5 kg (range -4.1 to 0) and the median volume of plasma treated per session was 5000 mL (range 4000 to 8600). In addition, the tandem procedure was as well tolerated as that for IA followed by HD., Conclusions: Tandem IA plus HD is a safe and cost-effective procedure.

Objectives: We implemented a desensitization program at our center to enable transplant in kidney-transplant candidates who have a living human-leukocyte antigen-incompatible (HLAi) donor. We report on the efficacy of semispecific immunoadsorption to allow HLAi kidney transplant in 6 highly sensitized patients., Materials and Methods: We chose immunoadsorption as the apheresis technique coupled to hemodialysis as a means to decrease donor-specific alloantibodies in kidney transplant candidates submitted to a pretransplant desensitization program to remove detrimental antibodies., Results: Six highly sensitized kidney-transplant patients (5 females), awaiting their first (n = 1) or second (n = 5) kidney transplant from a living donor, were enrolled in this desensitization program. They had 1 (n = 2), 2 (n = 1), 3 (n = 2), or 4 (n = 1) donor-specific alloantibodies; their mean fluorescent intensities at predesensitization ranged from 1200 to 19 000. Each patient underwent between 10 and 16 immunoadsorption sessions. At the time of transplant, donor-specific alloantibodies were undetectable in 2 patients (A24, DR3); donorspecific alloantibodies decreased by > 50% in 8 patients (A11, B44, DR3, DR11, DQ3 thrice, DQ5); donor-specific alloantibodies remained unchanged in 2 patients (B50, DR13); and mean fluorescent intensities were slightly increased in 2 patients (Cw6, DQ8). In the analysis of final outcomes, 2 patients experienced no rejection (1 experienced donor-specific alloantibody elimination, and 1 experienced a > 50% decrease in donor-specific alloantibodies). One patient presented with acute antibody-mediated rejection, which required immunoadsorption sessions and eculizumab therapy (donor-specific alloantibodies between 5000 and 19 000). Two patients presented with subacute antibody-mediated rejection; 1 was treated by plasmapheresis/rituximab therapy, and the other was treated with plasmapheresis/ methylprednisolone pulses. Another patient presented with chronic antibody-mediated rejection, which was treated unsuccessfully with plasmapheresis/rituximab; a tentative of rescue therapy with eculizumab was attempted without success., Conclusions: Desensitization of the humanleukocyte antigen using this immunoadsorption procedure effectively reduced or eliminated donorspecific alloantibodies in 71% of patients undergoing kidney transplant, at the time of transplant.

Objectives: We conducted a desensitization program in our center in patients undergoing kidney transplant for end-stage renal disease. These patients had a living-donor either ABO incompatible and/or human-leukocyte antigen-incompatible. The safety and efficacy of this program were evaluated., Materials and Methods: A pretransplant desensitization program relies on immunosuppressants and apheresis to remove detrimental antibodies. We chose immunoadsorption as the apheresis technique, and coupled this with hemodialysis in a tandem procedure., Results: We report on the efficacy of this new method in 120 procedures performed in 20 patients (14 ABO incompatible, 6 ABO incompatible/human leukocyte antigen-incompatible). The tandem procedure was well tolerated, and saved time compared with conducting sequential immunoadsorption and hemodialysis (6 h vs 10 h). The tandem procedure was associated with significantly decreased isoagglutinin titers and donor-specific alloantibodies (assessed by mean fluorescence intensity). Dialysance was effective (183, 102-264). The biochemical and hematologic parameters were similar to those observed after a conventional hemodialysis session, with the exception of protidemia; this might be related to some degree of albumin loss during the immunoadsoprtion procedure. The posttransplant events included 1) one ABO incompatible / human leukocyte antigenincompatible patient with vein thrombosis and ultimate kidney loss; 2) two patients with steroidsensitive cellular acute rejection; and 3) two patients with acute antibody-mediated rejection, which was successfully treated with apheresis and steroid pulses, plus rituximab in one and eculizumab in the other., Conclusions: We conclude that the tandem immunoadsorption-hemodialysis procedure is efficient at desensitizing patients with end-stage renal disease who are candidates for a living ABO incompatible and/or human leukocyte antigenincompatible donor-kidney transplant.

There is an urgent need for the development of alternative methods to replace animal testing for the prediction of repeat dose chemical toxicity. To address this need, the European Commission and Cosmetics Europe have jointly funded a research program for 'Safety Evaluation Ultimately Replacing Animal Testing.' The goal of this program was the development of in vitro cellular systems and associated computational capabilities for the prediction of hepatic, cardiac, renal, neuronal, muscle, and skin toxicities. An essential component of this effort is the choice of appropriate reference compounds that can be used in the development and validation of assays. In this review, we focus on the selection of reference compounds for liver pathologies in the broad categories of cytotoxicity and lipid disorders. Mitochondrial impairment, oxidative stress, and apoptosis are considered under the category of cytotoxicity, while steatosis, cholestasis, and phospholipidosis are considered under the category of lipid dysregulation. We focused on four compound classes capable of initiating such events, i.e., chemically reactive compounds, compounds with specific cellular targets, compounds that modulate lipid regulatory networks, and compounds that disrupt the plasma membrane. We describe the molecular mechanisms of these compounds and the cellular response networks which they elicit. This information will be helpful to both improve our understanding of mode of action and help in the selection of appropriate mechanistic biomarkers, allowing us to progress the development of animal-free models with improved predictivity to the human situation.

Background: Many kidney-transplant candidates have anti-HLA alloantibodies (HLAi): these make transplantation difficult, even from a living kidney (LK) donor, because of the presence of donor-specific anti-HLA alloantibodies. Due to the shortage of deceased kidney donors, the number of LK transplants is increasing, but is potentially limited by ABO incompatibility (ABOi)., Objectives: To make ABOi and/or HLAi patients suitable for kidney transplantation they need to be desensitised: this strategy is mainly based on rituximab therapy combined with either plasmapheresis (PP) or immunoadsorption (IA). IA can be more efficient than PP because greater plasma volumes can be treated within a single session than a PP session (>4 vs. 1.5-2). IA can be specific (ABOi setting) or non-specific (HLAi)., Design: We describe how we designed and implemented a desensitisation programme based on IA. This was started in the first trimester of 2010 within the Acute Polyvalent Haemodialysis and Apheresis Unit in Toulouse University Hospital, France. So far, we have performed >200 IA sessions with good results., Results: The IA sessions were associated with a net body-weight gain of ∼1 kg. Normally, IA is performed first and then haemodialysis on the same or next day; however, we have been able to, for the first time, couple IA with haemodialysis. Moreover, we can now carry out this procedure 24 hours a day, seven days a week., Conclusion: This procedure has improved patient care and reduced costs. The IA desensitisation programme has enabled successful transplantation in 24 patients to date., (© 2014 European Dialysis and Transplant Nurses Association/European Renal Care Association.)

Purpose: Evaluation of the psychometric properties of a cross-culturally adapted questionnaire, the Core Outcome Measurement Index for neck pain (COMI-neck)., Methods: The COMI-neck was cross-culturally adapted for the Italian language using established procedures. The following psychometric properties of the instrument were then assessed in patients with chronic neck pain undergoing rehabilitation: test-retest reliability (intraclass correlation coefficient, ICC); construct validity by comparing COMI-neck with the Neck Pain and Disability Scale, a numerical pain rating scale, and the EuroQol-Five Dimension (Pearson's correlations); and responsiveness by means of Standardized Response Mean (SRM), unpaired t tests, and Receiver Operating Characteristics (ROC) curves., Results: The questionnaire was completed by 103 subjects. The COMI-neck summary score displayed no relevant floor or ceiling effects. Test-retest reliability was excellent (ICC = 0.87). With one exception (symptom-specific well-being), the individual COMI items and the COMI summary score correlated to the expected extent with the scores of the reference questionnaires (r = 0.40-0.80). The mean change scores for the Italian COMI-neck differed significantly between patients with a good global outcome and those with a poor outcome (p = 0.002); SRM for the good outcome group was 1.23, and for the poor outcome group 0.40. ROC analysis revealed an area under the curve of 0.73 (95% CI: 0.62-0.85)., Conclusions: This study provides evidence that the Italian version of the COMI-neck is a valid and responsive questionnaire in the population of patients examined. Its use is recommended for clinical and research purposes.

Regularly monitoring blood flow through a vascular access (Qa) can predict a dysfunction and dramatically reduce the number of thromboses. The aim of our study was to compare two integrated access flow devices, thermodilution (Qa-BTM: BTM(®), Fresenius Medical Care, Bad Homburg, Germany) and ionic dialysance (Qa-ID: OCM(®), Fresenius Medical Care, Bad Homburg, Germany), with the "gold standard" saline dilution (Qa-T: Transonic(®), Systems Inc., Ithaca, NY, USA). Measurements were performed sequentially and were repeated in the first 90 minutes of a single dialysis session in 24 long-term hemodialysis patients with a vascular access. Bland-Altman, linear regression (r(2)), and intraclass correlation coefficients (ICC) assessed reproducibility, correlations, and concordance between the techniques. Average access flow for Qa-T was 1549 (± 844) mL/minute, Qa-BTM was 1530 (± 856) mL/minute (P = NS), and Qa-ID was 1619 (± 1085) mL/minute (P = NS). Respectively, ICC, (r(2)), and bias were 0.99, (0.98), and -19 mL/minute for Qa-BTM, and 0.75, (0.65), and +69 mL/minute for Qa-ID. The limits of agreement were -287 to +250 mL/minute for Qa-BTM and -1647 to +1785 mL/minute for Qa-ID. Reproducibility of thermodilution and ionic dialysance, expressed as relative differences, was not significantly different from saline dilution. Recirculation, measured by saline dilution, was 0% (0-4%), the same as the 0% measured by thermodilution, with correct placement of bloodlines and corrected for cardiopulmonary recirculation. The integrated access flow measurement devices, thermodilution and ionic dialysance, are reasonable alternatives to using saline dilution to measure Qa: Thermodilution showed better precision and correlation. They are reliable, make monitoring of vascular access easier, incur no extra costs, and use no additional consumables., (© 2013 International Society for Hemodialysis.)

We investigated contamination by endocrine-disrupting chemicals in drinking water from 35 major Italian cities and five popular Italian brands of bottled mineral water. The quality of Italian drinking water was assessed by combing chemical analysis with bioassay to quantify specific estrogenic contaminants and to characterize the actual biological effect of the mixture of chemicals present in drinking water including the contribution of not targeted compounds. The selected contaminants were natural and synthetic steroid estrogens, alkylphenols and bisphenol A, linuron, triazine herbicides, and their metabolites. A specific analytical method was developed based on solid phase extraction of 1 L of water and concentration to 100 μL for quantification by electrospray ionization liquid chromatography tandem mass spectrometry, achieving quantification limits of 0.05-0.36 ng/L for herbicides and 0.64-7.70 ng/L for steroids and phenols. No steroid estrogens were detected in any of the samples, while bisphenol A and nonylphenols were detected in the ranges of 0.82-102.00 and 10.30-84.00 ng/L respectively. Herbicides and their degradation products, when present, were found from slightly above the quantification limits up to 49.91 ng/L, mainly from cities in northern Italy. Chemical analyses were complemented by the performance of a bioassay for the determination of the estrogenic activity in the extracts based on the transactivation of estrogen receptor α-transfected reporter HeLa-ERE-Luciferase-Neomycin cell line. Activity was generally low with maximum estrogenicity of 13.6 pg/L estradiol equivalents.

The aim of the SEURAT-1 (Safety Evaluation Ultimately Replacing Animal Testing-1) research cluster, comprised of seven EU FP7 Health projects co-financed by Cosmetics Europe, is to generate a proof-of-concept to show how the latest technologies, systems toxicology and toxicogenomics can be combined to deliver a test replacement for repeated dose systemic toxicity testing on animals. The SEURAT-1 strategy is to adopt a mode-of-action framework to describe repeated dose toxicity, combining in vitro and in silico methods to derive predictions of in vivo toxicity responses. ToxBank is the cross-cluster infrastructure project whose activities include the development of a data warehouse to provide a web-accessible shared repository of research data and protocols, a physical compounds repository, reference or "gold compounds" for use across the cluster (available via wiki.toxbank.net), and a reference resource for biomaterials. Core technologies used in the data warehouse include the ISA-Tab universal data exchange format, REpresentational State Transfer (REST) web services, the W3C Resource Description Framework (RDF) and the OpenTox standards. We describe the design of the data warehouse based on cluster requirements, the implementation based on open standards, and finally the underlying concepts and initial results of a data analysis utilizing public data related to the gold compounds., (Copyright © 2013 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.)

A method based on ultrasonic extraction and purification by solid phase extraction followed by LC-MS/MS and GC-MS analysis was developed for the determination of genistein, genistin, iprodione, vinclozolin and procymidone in infant powdered formulas. The method was tested for different formulations: milk, soy and hypoallergenic, and was applied to European pooled samples. Spike recoveries ranged from 53.1% to 91.5% and the relative standard deviation values for repeatability ranged from 9.6% to 17.7%, except for iprodione in milk formula (22.3%). None of the fungicides were found in the European pooled formulae, while genistein was found at 9.7μg/g in soy formula and the concentration of genistin, its β-glycosylated form, was respectively 31.4ng/g and 476ng/g in milk and soy formula., (Copyright © 2012 Elsevier Ltd. All rights reserved.)

Solid wastes constitute an important and emerging problem. Landfills are still one of the most common ways to manage waste disposal. The risk assessment of pollutants from landfills is becoming a major environmental issue in Europe, due to the large number of sites and to the importance of groundwater protection. Furthermore, there is lack of knowledge for the environmental, ecotoxicological and toxicological characteristics of most contaminants contained into landfill leacheates. Understanding leachate composition and creating an integrated strategy for risk assessment are currently needed to correctly face the landfill issues and to make projections on the long-term impacts of a landfill, with particular attention to the estimation of possible adverse effects on human health and ecosystem. In the present study, we propose an integrated strategy to evaluate the toxicity of the leachate using chemical analyses, risk assessment guidelines and in vitro assays using the hepatoma HepG2 cells as a model. The approach was applied on a real case study: an industrial waste landfill in northern Italy for which data on the presence of leachate contaminants are available from the last 11 years. Results from our ecological risk models suggest important toxic effects on freshwater fish and small rodents, mainly due to ammonia and inorganic constituents. Our results from in vitro data show an inhibition of cell proliferation by leachate at low doses and cytotoxic effect at high doses after 48 h of exposure., (Copyright © 2011 Elsevier Inc. All rights reserved.)

This paper reports the results obtained from the data collected within the European Commission funded project SAFEFOODNET regarding the state of the art in the control of chemical food contaminants in twelve European New Member States and one Associated Candidate Country (Turkey). Information has been gathered on institutions involved in food chemical contamination control, types of contaminants and matrices analyzed, procedures for data quality assurance, purposes of the analyses and accessibility of data in the participant countries. The resulting picture points out the general availability of adequate capabilities for the analysis of food contaminants in the laboratories in charge of control and the performance of the analysis of a large variety of chemicals (persistent organic pollutants, polycyclic aromatic hydrocarbons, pesticides, mycotoxins, heavy metals, radionuclides) in almost each country with few exceptions (dioxins in Bulgaria, Turkey, Latvia, persistent organic pollutants in Lithuania and Malta, polycyclic aromatic hydrocarbons in Malta). The application of validated analytical methods and the process of laboratory accreditation are partially fulfilled within the investigated countries, but still forthcoming for some countries, as in Romania, Turkey and Malta. Information collected on food controls is only partially available online and the language used is prevalently local and English to a lesser extent.

An effects-directed strategy was applied to bed sediments of a polluted tributary in order to isolate and identify the major estrogenic chemicals it discharges into the River Po, the principal Italian watercourse. Sediment extract was concentrated by solid phase extraction and then fractioned into 10 fractions by reversed phase high performance liquid chromatography (RP-HPLC). Estrogenic activity of whole extract and fractions were determined using a recombinant yeast assay containing the human estrogen receptor (YES). The 10 fractions and whole extract were analysed for target compounds, e.g. estrone (E1), 17beta-estradiol (E2), estriol (E3), 4-nonylphenol (NP), 4-tert-octylphenol (t-OP), bisphenol A (BPA), using both liquid chromatography-tandem mass spectrometry (LC-MS/MS) and non-competitive enzyme-linked immunosorbent assays (ELISA). The YES assay determined high estrogenic activity in whole sediment (15.6 ng/g EE2 equivalents), and positive results for fractions nr 1, 2, 6, 7 and 8. E1, E3 and NP were the main estrogenic chemicals, however, other unidentified compounds contributed to sediment estrogenicity, particularly for polar fractions nr 1 and 2. A GC-MS screening performed in scan mode identified other potential contributors such as phthalates (DBP, BBP), and OP isomers. A next sampling campaign extended to other tributaries and receiving stretches of the River Po confirmed E1, E3 and NP as major estrogenic chemicals potentially threatening other sites of the main river. In general, target compound ELISAs have been shown to be suitable tools for a rapid screening of wide areas or large numbers of environmental samples for estrogenic risk. The potential for interferences suggests however to use cautiously the concentration values obtained from some of the immunoassays.