A Case Study of Upper Limb Robotic-Assisted Therapy Using the Track-Hold Device.

The Track-Hold System (THS) project, developed in a healthcare facility and therefore in a controlled and protected healthcare environment, contributes to the more general and broad context of Robotic-Assisted Therapy (RAT). RAT represents an advanced and innovative rehabilitation method, both motor and cognitive, and uses active, passive, and facilitating robotic devices. RAT devices can be equipped with sensors to detect and track voluntary and involuntary movements. They can work in synergy with multimedia protocols developed ad hoc to achieve the highest possible level of functional re-education. The THS is based on a passive robotic arm capable of recording and facilitating the movements of the upper limbs. An operational interface completes the device for its use in the clinical setting. In the form of a case study, the researchers conducted the experimentation in the former Tabarracci hospital (Viareggio, Italy). The case study develops a motor and cognitive rehabilitation protocol. The chosen subjects suffered from post-stroke outcomes affecting the right upper limb, including strength deficits, tremors, incoordination, and motor apraxia. During the first stage of the enrolment, the researchers worked with seven patients. The researchers completed the pilot with four patients because three of them got a stroke recurrence. The collaboration with four patients permitted the generation of an enlarged case report to collect preliminary data. The preliminary clinical results of the Track-Hold System Project demonstrated good compliance by patients with robotic-assisted rehabilitation; in particular, patients underwent a gradual path of functional recovery of the upper limb using the implemented interface.

A System for Neuromotor Based Rehabilitation on a Passive Robotic Aid.

In the aging world population, the occurrence of neuromotor deficits arising from stroke and other medical conditions is expected to grow, demanding the design of new and more effective approaches to rehabilitation. In this paper, we show how the combination of robotic technologies with progress in exergaming methodologies may lead to the creation of new rehabilitation protocols favoring motor re-learning. To this end, we introduce the Track-Hold system for neuromotor rehabilitation based on a passive robotic arm and integrated software. A special configuration of weights on the robotic arm fully balances the weight of the patients' arm, allowing them to perform a purely neurological task, overcoming the muscular effort of similar free-hand exercises. A set of adaptive and configurable exercises are proposed to patients through a large display and a graphical user interface. Common everyday tasks are also proposed for patients to learn again the associated actions in a persistent way, thus improving life independence. A data analysis module was also designed to monitor progress and compute indices of post-stroke neurological damage and Parkinsonian-type disorders. The system was tested in the lab and in a pilot project involving five patients in the post-stroke chronic stage with partial paralysis of the right upper limb, showing encouraging preliminary results.

Quantification of Tumor Vasculature by Analysis of Amount and Spatial Dispersion of Caliber-Classified Vessels.

This protocol focuses on the quantitative description of the angioarchitecture of experimental tumor xenografts. This semiautomatic analysis is carried out on functional vessels and microvessels acquired by confocal imaging and processed into progressively reconstructed angioarchitectures following a caliber-classification step. The protocol can be applied also to the quantification of pathological angioarchitectures other than tumor grafts as well as to the microvasculature of physiological tissue samples.

Quantification of 3D Brain Microangioarchitectures in an Animal Model of Krabbe Disease.

We performed a three-dimensional (3D) analysis of the microvascular network of the cerebral cortex of twitcher mice (an authentic model of Krabbe disease) using a restricted set of indexes that are able to describe the arrangement of the microvascular tree in CD31-stained sections. We obtained a near-linear graphical "fingerprint" of the microangioarchitecture of wild-type and twitcher animals that describes the amounts, spatial dispersion, and spatial relationships of adjacent classes of caliber-filtered microvessels. We observed significant alterations of the microangioarchitecture of the cerebral cortex of twitcher mice, whereas no alterations occur in renal microvessels, which is keeping with the observation that kidney is an organ that is not affected by the disease. This approach may represent an important starting point for the study of the microvascular changes that occur in the central nervous system (CNS) under different physiopathological conditions.

ß-Galactosylceramidase Deficiency Causes Bone Marrow Vascular Defects in an Animal Model of Krabbe Disease.

Krabbe disease (KD) is an autosomal recessive sphingolipidosis caused by the deficiency of the lysosomal hydrolase ß-galactosylceramidase (GALC). Oligodendroglia degeneration and demyelination of the nervous system lead to neurological dysfunctions which are usually lethal by two years of age. At present, the only clinical treatment with any proven efficacy is hematopoietic stem-cell transplantation, which is more effective when administered in the neonatal period to presymptomatic recipients. Bone marrow (BM) sinusoidal endothelial cells (SECs) play a pivotal role in stem cell engraftment and reconstitution of hematopoiesis. Previous observations had shown significant alterations of microvascular endothelial cells in the brain of KD patients and in Galc mutant twitcher mice, an authentic model of the disease. In the present study, we investigated the vascular component of the BM in the femurs of symptomatic homozygous twitcher mice at postnatal day P36. Histological, immunohistochemical, and two-photon microscopy imaging analyses revealed the presence of significant alterations of the diaphyseal BM vasculature, characterized by enlarged, discontinuous, and hemorrhagic SECs that express the endothelial marker vascular endothelial growth factor receptor-2 (VEGFR2) but lack platelet/endothelial cell adhesion molecule-1 (CD31) expression. In addition, computer-aided image analysis indicates that twitcher CD31-/VEGFR2+ SECs show a significant increase in lumen size and in the number and size of endothelial gaps compared to BM SECs of wild type littermates. These results suggest that morphofunctional defects in the BM vascular niche may contribute to the limited therapeutic efficacy of hematopoietic stem-cell transplantation in KD patients at symptomatic stages of the disease.

Vascular amounts and dispersion of caliber-classified vessels as key parameters to quantitate 3D micro-angioarchitectures in multiple myeloma experimental tumors.

Blood vessel micro-angioarchitecture plays a pivotal role in tumor progression, metastatic dissemination and response to therapy. Thus, methods able to quantify microvascular trees and their anomalies may allow a better comprehension of the neovascularization process and evaluation of vascular-targeted therapies in cancer. To this aim, the development of a restricted set of indexes able to describe the arrangement of a microvascular tree is eagerly required. We addressed this goal through 3D analysis of the functional microvascular network in sulfo-biotin-stained human multiple myeloma KMS-11 xenografts in NOD/SCID mice. Using image analysis, we show that amounts, spatial dispersion and spatial relationships of adjacent classes of caliber-filtered microvessels provide a near-linear graphical "fingerprint" of tumor micro-angioarchitecture. Position, slope and axial projections of this graphical outcome reflect biological features and summarize the properties of tumor micro-angioarchitecture. Notably, treatment of KMS-11 xenografts with anti-angiogenic drugs affected position and slope of the specific curves without degrading their near-linear properties. The possibility offered by this procedure to describe and quantify the 3D features of the tumor micro-angioarchitecture paves the way to the analysis of the microvascular tree in human tumor specimens at different stages of tumor progression and after pharmacologic interventions, with possible diagnostic and prognostic implications.

The fifth subunit in α3ß4 nicotinic receptor is more than an accessory subunit.

The α3ß4 subtype is the predominant neuronal nicotinic acetylcholine receptor present in the sensory and autonomic ganglia and in a subpopulation of brain neurons. This subtype can form pentameric receptors with either 2 or 3 ß4 subunits that have different pharmacologic and functional properties. To further investigate the role of the fifth subunit, we coexpressed a dimeric construct coding for a single polypeptide containing the ß4 and α3 subunit sequences, with different monomeric subunits. With this strategy, which allowed the formation of single populations of receptors with unique stoichiometry, we demonstrated with immunofluorescence and biochemical and functional assays that only the receptors with 3 ß4 subunits are efficiently expressed at the plasma membrane. Moreover, the LFM export motif of ß4 subunit in the fifth position exerts a unique function in the regulation of the intracellular trafficking of the receptors, their exposure at the cell surface, and consequently, their function, whereas the same export motif present in the ß4 subunits forming the acetylcholine binding site is dispensable.-Crespi, A., Plutino, S., Sciaccaluga, M., Righi, M., Borgese, N., Fucile, S., Gotti, C., Colombo, S. F. The fifth subunit in α3ß4 nicotinic receptor is more than an accessory subunit.

Brain angioarchitecture and intussusceptive microvascular growth in a murine model of Krabbe disease.

Defects of the angiogenic process occur in the brain of twitcher mouse, an authentic model of human Krabbe disease caused by genetic deficiency of lysosomal ß-galactosylceramidase (GALC), leading to lethal neurological dysfunctions and accumulation of neurotoxic psychosine in the central nervous system. Here, quantitative computational analysis was used to explore the alterations of brain angioarchitecture in twitcher mice. To this aim, customized ImageJ routines were used to assess calibers, amounts, lengths and spatial dispersion of CD31(+) vessels in 3D volumes from the postnatal frontal cortex of twitcher animals. The results showed a decrease in CD31 immunoreactivity in twitcher brain with a marked reduction in total vessel lengths coupled with increased vessel fragmentation. No significant changes were instead observed for the spatial dispersion of brain vessels throughout volumes or in vascular calibers. Notably, no CD31(+) vessel changes were detected in twitcher kidneys in which psychosine accumulates at very low levels, thus confirming the specificity of the effect. Microvascular corrosion casting followed by scanning electron microscopy morphometry confirmed the presence of significant alterations of the functional angioarchitecture of the brain cortex of twitcher mice with reduction in microvascular density, vascular branch remodeling and intussusceptive angiogenesis. Intussusceptive microvascular growth, confirmed by histological analysis, was paralleled by alterations of the expression of intussusception-related genes in twitcher brain. Our data support the hypothesis that a marked decrease in vascular development concurs to the onset of neuropathological lesions in twitcher brain and suggest that neuroinflammation-driven intussusceptive responses may represent an attempt to compensate impaired sprouting angiogenesis.

A wize mirror for lifestyle improvement.

This paper discusses the problem of fostering lifestyle changes towards healthier habits via tailored user guidance. We present a novel multisensory device, the Wize Mirror, which will be able to detect semeiotic face signs related to cardio-metabolic risk, and encourage users to reduce their risk by improving their lifestyle. Offering a proper user guidance requires solving three main issues: user profiling, definition of a wellness index based on biophysical data, and personalized guidance by means of coaching and supportive messages. For each of these issues, the solutions proposed in the EU FP7 Project SEMEOTICONS are presented, highlighting their advantages with respect to the state-of-the-art.