Development of engineered nanoparticles for biomedical and industrial applications.

In this work four nanoparticle systems were designed: two for medical and two for industrial applications. The purpose of the first part of my study concerns the manufacture of a NPs-enzyme system for cancer therapy: Fe3O4 NPs@APTES-DAAO. This system combine the magnetic properties of iron oxide nanoparticles (Fe3O4 NPs) with the ROS generating enzyme D-amino acid oxidase (DAAO) in order to efficiently direct the enzyme into the tumor and kill tumor cells. The second medical system designed consists in magnetic iron oxide nanoparticles conjugated with the antibiotics teicoplanin. Similar to the previous system, Fe3O4 NPs@Teicoplanin wants to takes advantage of the magnetic character of NPs for a specific targeting of the antibiotic. The two systems for industrial application were composed of Fe3O4 NPs conjugated to a cephalosporin C acylase (VAC) and a L-aspartate oxidase (LASPO) respectively. In industrial enzymatic application processes, immobilization of enzymes can offer several advantages, including the ability to be used repeatedly, improvement of enzyme stability and broadening the optimum pH range of enzyme. The first system Fe3O4 NPs@APTES-VAC allows the one-step conversion of cephalosporin C in 7-aminocephalosporanic acid, which is the precursor of in semisynthetic cephalosporins antibiotics. The second system Fe3O4 NPs@APTES-LASPO allows the resolution of a racemic mixture of D, L-aspartate

Development of engineered nanoparticles for biomedical and industrial applications.

In this work four nanoparticle systems were designed: two for medical and two for industrial applications. The purpose of the first part of my study concerns the manufacture of a NPs-enzyme system for cancer therapy: Fe3O4 NPs@APTES-DAAO. This system combine the magnetic properties of iron oxide nanoparticles (Fe3O4 NPs) with the ROS generating enzyme D-amino acid oxidase (DAAO) in order to efficiently direct the enzyme into the tumor and kill tumor cells. The second medical system designed consists in magnetic iron oxide nanoparticles conjugated with the antibiotics teicoplanin. Similar to the previous system, Fe3O4 NPs@Teicoplanin wants to takes advantage of the magnetic character of NPs for a specific targeting of the antibiotic. The two systems for industrial application were composed of Fe3O4 NPs conjugated to a cephalosporin C acylase (VAC) and a L-aspartate oxidase (LASPO) respectively. In industrial enzymatic application processes, immobilization of enzymes can offer several advantages, including the ability to be used repeatedly, improvement of enzyme stability and broadening the optimum pH range of enzyme. The first system Fe3O4 NPs@APTES-VAC allows the one-step conversion of cephalosporin C in 7-aminocephalosporanic acid, which is the precursor of in semisynthetic cephalosporins antibiotics. The second system Fe3O4 NPs@APTES-LASPO allows the resolution of a racemic mixture of D, L-aspartate

Development of engineered nanoparticles for biomedical and industrial applications.

In this work four nanoparticle systems were designed: two for medical and two for industrial applications. The purpose of the first part of my study concerns the manufacture of a NPs-enzyme system for cancer therapy: Fe3O4 NPs@APTES-DAAO. This system combine the magnetic properties of iron oxide nanoparticles (Fe3O4 NPs) with the ROS generating enzyme D-amino acid oxidase (DAAO) in order to efficiently direct the enzyme into the tumor and kill tumor cells. The second medical system designed consists in magnetic iron oxide nanoparticles conjugated with the antibiotics teicoplanin. Similar to the previous system, Fe3O4 NPs@Teicoplanin wants to takes advantage of the magnetic character of NPs for a specific targeting of the antibiotic. The two systems for industrial application were composed of Fe3O4 NPs conjugated to a cephalosporin C acylase (VAC) and a L-aspartate oxidase (LASPO) respectively. In industrial enzymatic application processes, immobilization of enzymes can offer several advantages, including the ability to be used repeatedly, improvement of enzyme stability and broadening the optimum pH range of enzyme. The first system Fe3O4 NPs@APTES-VAC allows the one-step conversion of cephalosporin C in 7-aminocephalosporanic acid, which is the precursor of in semisynthetic cephalosporins antibiotics. The second system Fe3O4 NPs@APTES-LASPO allows the resolution of a racemic mixture of D, L-aspartate

Clinical Relevance of State-of-the-Art Analysis of Surface Electromyography in Cerebral Palsy

Surface electromyography (sEMG) can be used to assess the integrity of the neuromuscular system and its impairment in neurological disorders. Here we will consider several issues related to the current clinical applications, difficulties and limited usage of sEMG for the assessment and rehabilitation of children with cerebral palsy. The uniqueness of this methodology is that it can determine hyperactivity or inactivity of selected muscles, which cannot be assessed by other methods. In addition, it can assist for intervention or muscle/tendon surgery acts, and it can evaluate integrated functioning of the nervous system based on multi-muscle sEMG recordings and assess motor pool activation. The latter aspect is especially important for understanding impairments of the mechanisms of neural controllers rather than malfunction of individual muscles. Although sEMG study is an important tool in both clinical research and neurorehabilitation, the results of a survey on the clinical relevance of sEMG in a typical department of pediatric rehabilitation highlighted its limited clinical usage. We believe that this is due to limited knowledge of the sEMG and its neuromuscular underpinnings by many physiotherapists, as a result of lack of emphasis on this important methodology in the courses taught in physical therapy schools. The lack of reference databases or benchmarking software for sEMG analysis may also contribute to the limited clinical usage. Despite the existence of educational and technical barriers to a widespread use of, sEMG does provide important tools for planning and assessment of rehabilitation treatments for children with cerebral palsy

Evaluation of Spatiotemporal Patterns of the Spinal Muscle Coordination Output during Walking in the Exoskeleton

Recent advances in the performance and evaluation of walking in exoskeletons use various assessments based on kinematic/kinetic measurements. While such variables provide general characteristics of gait performance, only limited conclusions can be made about the neural control strategies. Moreover, some kinematic or kinetic parameters are a consequence of the control implemented on the exoskeleton. Therefore, standard indicators based on kinematic variables have limitations and need to be complemented by performance measures of muscle coordination and control strategy. Knowledge about what happens at the spinal cord output level might also be critical for rehabilitation since an abnormal spatiotemporal integration of activity in specific spinal segments may result in a risk for abnormalities in gait recovery. Here we present the PEPATO software, which is a benchmarking solution to assess changes in the spinal locomotor output during walking in the exoskeleton with respect to reference data on normal walking. In particular, functional and structural changes at the spinal cord level can be mapped into muscle synergies and spinal maps of motoneuron activity. A user-friendly software interface guides the user through several data processing steps leading to a set of performance indicators as output. We present an example of the usage of this software for evaluating walking in an unloading exoskeleton that allows a person to step in simulated reduced (the Moon's) gravity. By analyzing the EMG activity from lower limb muscles, the algorithms detected several performance indicators demonstrating differential adaptation (shifts in the center of activity, prolonged activation) of specific muscle activation modules and spinal motor pools and increased coactivation of lumbar and sacral segments. The software is integrated at EUROBENCH facilities to benchmark the performance of walking in the exoskeleton from the point of view of changes in the spinal locomotor output

Zerovalent Fe, Co and Ni nanoparticle toxicity evaluated on SKOV-3 and U87 cell lines

ABSTRACT:We have considered nanoparticles (NPs) of Fe, Co and Ni, three transition metals sharing similar chemical properties. NP dissolution, conducted by radioactive tracer method and inductively coupled plasmamass spectrometry, indicated that NiNPs and FeNPs released in the medium a much smaller amount of ions than that released by Co NPs. The two considered methodological approaches, however, gave comparable but not identical results. All NPs are readily internalized by the cells, but their quantity inside the cells is less than 5%. Cytotoxicity and gene expression experimentswere performed on SKOV-3 and U87 cells. In both cell lines, CoNPs and NiNPs were definitely more toxic than FeNPs. Real-time polymerase chain reaction experiments aimed to evaluatemodifications of the expression of genes involved in the cellular stress response (HSP70, MT2A), or susceptible to metal exposure (SDHB1 and MLL), or involved in specific cellular processes (caspase3, IQSEC1 and VMP1), gave different response patterns in the two cell lines. HSP70, for example, was highly upregulated by CoNPs and NiNPs, but only in SKOV-3 cell lines. Overall, this work underlines the difficulties in predicting NP toxicological properties based only on their chemical characteristics. We, consequently, think that, at this stage of our knowledge, biological effects induced by metal-based NPs should be examined on a case-by-case basis following studies on different in vitro models. Moreover, with the only exception of U87 exposed to Ni, our results suggest thatmetallic NPs have caused, on gene expression, similar effects to those caused by their cor- Q2 responding ions

A new severity score index for phenotypic classification and evaluation of responses to treatment in type I Gaucher disease

Gaucher disease is the first lysosomal storage disease for which specific therapy became available. Over 4800 patients have been treated with enzyme replacement therapy. Analysis of Gaucher disease registry data has outlined the clinical heterogeneity of the disease and the different responses to treatment from patient to patient, and for different organs. This variability in clinical response justifies the development of a severity score index to assess disease activity, stage and prognosis, and to quantify the effects of treatment.The \u201cGaucher Disease Severity Score Index Type I\u201d(GauSSI-I), is based on the clinical experience of the authors and an extensive literature review, including data from the International Gaucher Registry. In particular for skeletal disease, all the available scoring systems have been reviewed and compared in order to provide a skeletal scoring system that allows use of any of the different methods. Siix specific domains, in which different items were scored according to their impact on morbidity, were characterized. GauSSI-I was evaluated in 53 type I Gaucher patients treated with imiglucerase, and it was compared to the Zimran score, the only severity index score so far available. It is a reliable method for staging the severity of adult type I Gaucher disease, and it is more sensitive than the Zimran score for monitoring the response to treatment