by Keyword: AD

By year:[ 2020 | 2019 | 2018 | 2017 | 2016 | 2015 | 2014 | 2013 | 2012 | 2011 | 2010 | 2009 | 2008 | 2007 | 2006 | 2005 ]

Ferrer, Isidro, Andrés-Benito, Pol, Zelaya, Maria Victoria, Aguirre, Maria Elena Erro, Carmona, Margarita, Ausín, Karina, Lachén-Montes, Mercedes, Fernández-Irigoyen, Joaquín, Santamaría, Enrique, del Río, José Antonio, (2020). Familial globular glial tauopathy linked to MAPT mutations: molecular neuropathology and seeding capacity of a prototypical mixed neuronal and glial tauopathy Acta Neuropathologica 139, (4), 735-771

Globular glial tauopathy (GGT) is a progressive neurodegenerative disease involving the grey matter and white matter (WM) and characterized by neuronal deposition of hyper-phosphorylated, abnormally conformed, truncated, oligomeric 4Rtau in neurons and in glial cells forming typical globular astrocyte and oligodendrocyte inclusions (GAIs and GOIs, respectively) and coiled bodies. Present studies centre on four genetic GGT cases from two unrelated families bearing the P301T mutation in MAPT and one case of sporadic GGT (sGGT) and one case of GGT linked to MAPT K317M mutation, for comparative purposes. Clinical and neuropathological manifestations and biochemical profiles of phospho-tau are subjected to individual variations in patients carrying the same mutation, even in carriers of the same family, independently of the age of onset, gender, and duration of the disease. Immunohistochemistry, western blotting, transcriptomic, proteomics and phosphoproteomics, and intra-cerebral inoculation of brain homogenates to wild-type (WT) mice were the methods employed. In GGT cases linked to MAPT P301T mutation, astrocyte markers GFAP, ALDH1L1, YKL40 mRNA and protein, GJA1 mRNA, and AQ4 protein are significantly increased; glutamate transporter GLT1 (EAAT2) and glucose transporter (SLC2A1) decreased; mitochondrial pyruvate carrier 1 (MPC1) increased, and mitochondrial uncoupling protein 5 (UCP5) almost absent in GAIs in frontal cortex (FC). Expression of oligodendrocyte markers OLIG1 and OLIG2mRNA, and myelin-related genes MBP, PLP1, CNP, MAG, MAL, MOG, and MOBP are significantly decreased in WM; CNPase, PLP1, and MBP antibodies reveal reduction and disruption of myelinated fibres; and SMI31 antibodies mark axonal damage in the WM. Altered expression of AQ4, GLUC-t, and GLT-1 is also observed in sGGT and in GGT linked to MAPT K317M mutation. These alterations point to primary astrogliopathy and oligodendrogliopathy in GGT. In addition, GGT linked to MAPT P301T mutation proteotypes unveil a proteostatic imbalance due to widespread (phospho)proteomic dearrangement in the FC and WM, triggering a disruption of neuron projection morphogenesis and synaptic transmission. Identification of hyper-phosphorylation of variegated proteins calls into question the concept of phospho-tau-only alteration in the pathogenesis of GGT. Finally, unilateral inoculation of sarkosyl-insoluble fractions of GGT homogenates from GGT linked to MAPT P301T, sGGT, and GGT linked to MAPT K317M mutation in the hippocampus, corpus callosum, or caudate/putamen in wild-type mice produces seeding, and time- and region-dependent spreading of phosphorylated, non-oligomeric, and non-truncated 4Rtau and 3Rtau, without GAIs and GOIs but only of coiled bodies. These experiments prove that host tau strains are important in the modulation of cellular vulnerability and phenotypes of phospho-tau aggregates.

Keywords: Globular glial tauopathy, Tau, Astrogliopathy, Oligodendrogliopathy, Phosphoproteome, Seeding and spreading

Altay, Gizem, Batlle, Eduard, Fernández-Majada, Vanesa, Martínez, Elena, (2020). In vitro self-organized mouse small intestinal epithelial monolayer protocol Bio-protocol 10, (3), e3514

Developing protocols to obtain intestinal epithelial monolayers that recapitulate in vivo physiology to overcome the limitations of the organoids’ closed geometry has become of great interest during the last few years. Most of the developed culture models showed physiological-relevant cell composition but did not prove self-renewing capacities. Here, we show a simple method to obtain mouse small intestine-derived epithelial monolayers organized into proliferative crypt-like domains, containing stem cells, and differentiated villus-like regions, closely resembling the in vivo cell composition and distribution. In addition, we adapted our model to a tissue culture format compatible with functional studies and prove close to physiological barrier properties of our in vitro epithelial monolayers. Thus, we have set-up a protocol to generate physiologically relevant intestinal epithelial monolayers to be employed in assays where independent access to both luminal and basolateral compartments is needed, such as drug absorption, intracellular trafficking and microbiome-epithelium interaction assays.

Keywords: Mouse intestinal organoids, Adult intestinal stem cells, Matrigel, Intestinal epithelial monolayer, In vitro intestinal epithelial model, Tissue-like functionality, TEER

del Rio, Jose A., Ferrer, Isidre, (2020). Potential of microfluidics and lab-on-chip platforms to improve understanding of “prion-like” protein assembly and behavior Frontiers in Bioengineering and Biotechnology 8, 570692

Human aging is accompanied by a relevant increase in age-associated chronic pathologies, including neurodegenerative and metabolic diseases. The appearance and evolution of numerous neurodegenerative diseases is paralleled by the appearance of intracellular and extracellular accumulation of misfolded proteins in affected brains. In addition, recent evidence suggests that most of these amyloid proteins can behave and propagate among neural cells similarly to infective prions. In order to improve understanding of the seeding and spreading processes of these “prion-like” amyloids, microfluidics and 3D lab-on-chip approaches have been developed as highly valuable tools. These techniques allow us to monitor changes in cellular and molecular processes responsible for amyloid seeding and cell spreading and their parallel effects in neural physiology. Their compatibility with new optical and biochemical techniques and their relative availability have increased interest in them and in their use in numerous laboratories. In addition, recent advances in stem cell research in combination with microfluidic platforms have opened new humanized in vitro models for myriad neurodegenerative diseases affecting different cellular targets of the vascular, muscular, and nervous systems, and glial cells. These new platforms help reduce the use of animal experimentation. They are more reproducible and represent a potential alternative to classical approaches to understanding neurodegeneration. In this review, we summarize recent progress in neurobiological research in “prion-like” protein using microfluidic and 3D lab-on-chip approaches. These approaches are driven by various fields, including chemistry, biochemistry, and cell biology, and they serve to facilitate the development of more precise human brain models for basic mechanistic studies of cell-to-cell interactions and drug discovery.

Keywords: Lab-On-Chip, Amyloid propagation, Microfluidics, Fibril, Seeding, Spreading, Prion-like, Prionoid

Ferrer, I., Zelaya, M. V., Aguiló García, M., Carmona, M., López-González, I., Andrés-Benito, P., Lidón, L., Gavín, R., Garcia-Esparcia, P., del Rio, J. A., (2020). Relevance of host tau in tau seeding and spreading in tauopathies Brain Pathology 30, (2), 298-318

Human tau seeding and spreading occur following intracerebral inoculation of brain homogenates obtained from tauopathies in transgenic mice expressing natural or mutant tau, and in wild-type (WT) mice. The present study was geared to learning about the patterns of tau seeding, the cells involved and the characteristics of tau following intracerebral inoculation of homogenates from primary age-related tauopathy (PART: neuronal 4Rtau and 3Rtau), aging-related tau astrogliopathy (ARTAG: astrocytic 4Rtau) and globular glial tauopathy (GGT: 4Rtau with neuronal deposits and specific tau inclusions in astrocytes and oligodendrocytes). For this purpose, young and adult WT mice were inoculated unilaterally in the hippocampus or in the lateral corpus callosum with sarkosyl-insoluble fractions from PART, ARTAG and GGT cases, and were killed at variable periods of three to seven months. Brains were processed for immunohistochemistry in paraffin sections. Tau seeding occurred in the ipsilateral hippocampus and corpus callosum and spread to the septal nuclei, periventricular hypothalamus and contralateral corpus callosum, respectively. Tau deposits were mainly found in neurons, oligodendrocytes and threads; the deposits were diffuse or granular, composed of phosphorylated tau, tau with abnormal conformation and 3Rtau and 4Rtau independently of the type of tauopathy. Truncated tau at the aspartic acid 421 and ubiquitination were absent. Tau deposits had the characteristics of pre-tangles. A percentage of intracellular tau deposits co-localized with active (phosphorylated) tau kinases p38 and ERK 1/2. Present study shows that seeding and spreading of human tau into the brain of WT mice involves neurons and glial cells, mainly oligodendrocytes, thereby supporting the idea of a primary role of oligodendrogliopathy, together with neuronopathy, in the progression of tauopathies. In addition, it suggests that human tau inoculation modifies murine tau metabolism with the production and deposition of 3Rtau and 4Rtau, and by activation of specific tau kinases in affected cells.

Keywords: Aging-related tau astrogliopathy, Globular glial tauopathy, Primary age-related tauopathy, Seeding, Spreading, Tau, Tauopathies

Kaang, Byung Kwon, Mestre, Rafael, Kang, Dong-Chang, Sánchez, Samuel, Kim, Dong-Pyo, (2020). Scalable and integrated flow synthesis of triple-responsive nano-motors via microfluidic Pickering emulsification Applied Materials Today 21, 100854

Artificial micro-/nano-motors are tiny machines as newly emerging tools capable of achieving numerous tasks. In principle, the self-phoretic motions require asymmetric structures in geometry and chemistry. However, conventional production techniques suffered from complex and time consuming multi-step process in low uniformity, and difficult to endow multi-functions into motors. This work disclosed a continuous-flow synthesis of triple-responsive (thermophoretic, chemical and magnetic movement) nano-motors (m-SiO2/Fe3O4-Pdop/Pt) via microfluidic Pickering emulsification in a process of integrated and scalable manner. The droplet microfluidic process allows efficient self-assembly of the silica nanoparticles surrounding the spherical interface of resin droplet, rendering excellent Pickering efficiency and reproducibility, and followed by anisotropic decoration of polydopamine (Pdop) and Pt catalyst in a serial flow process. The obtained Janus nanoparticles reveal double- or triple-responsive self-propulsions with synergic mobility by combining thermophoresis powered by light, catalytic driven motion in H2O2 or magnetic movement by magnet. Further, a non-metallic polydopamine based thermophoretic motion as well as an automated nano-cleaner for rapid water purification by dye removal are convincingly functioned. Finally, this novel integrated flow strategy proves a scalable manufacturing production (> 0.7 g hr−1) of the nano-motors using inexpensive single microreactor, fulfilling quantitative and qualitative needs for versatile applications.

Keywords: Microfluidics Pickering emulsions, Triple-responsive motor, Adsorbent

Solà-Soler, J., Giraldo, B. F., (2020). Comparison of ECG-eerived respiration estimation methods on healthy subjects in function of recording site and subject position and gender Engineering in Medicine & Biology Society (EMBC) 42nd Annual International Conference of the IEEE , IEEE (Montreal, Canada) , 2650-2653

Respiration rate can be assessed by analyzing respiratory changes of the electrocardiogram (ECG). Several methods can be applied to derive the respiratory signal from the ECG (EDR signal). In this study, four EDR estimation methods based on QRS features were analyzed. A database with 44 healthy subjects (16 females) in supine and sitting positions was analyzed. Respiratory flow and ECG recordings on leads I, II, III and a Chest lead was studied. A QR slope-based method, an RS slope-based method, an QRS angle-based method and an QRS area-based method were applied. Their performance was evaluated by the correlation coefficient with the reference respiratory volume signal. Significantly higher correlation coefficients in the range r = 0.77 – 0.86 were obtained with the Chest lead for all methods. The EDR estimation method based on the QRS angle provided the highest similarity with the volume signal for all recording leads and subject positions. We found no statistically significant differences according to gender or subject position.Clinical Relevance— This work analyzes the EDR signal from four electrocardiographic leads to obtain the respiratory signal and contributes to a simplified analysis of respiratory activity.

Keywords: Electrocardiography, Lead, Estimation, Correlation coefficient, Databases, Heart, Correlation

Lozano-García, M., Nuhic, J., Moxham, J., Rafferty, G. F., Jolley, C. J., Jané, R., (2020). Performance evaluation of fixed sample entropy for lung sound intensity estimation Engineering in Medicine & Biology Society (EMBC) 42nd Annual International Conference of the IEEE , IEEE (Montreal, Canada) , 2740-2743

Lung sound (LS) signals are often contaminated by impulsive artifacts that complicate the estimation of lung sound intensity (LSI) using conventional amplitude estimators. Fixed sample entropy (fSampEn) has proven to be robust to cardiac artifacts in myographic respiratory signals. Similarly, fSampEn is expected to be robust to artifacts in LS signals, thus providing accurate LSI estimates. However, the choice of fSampEn parameters depends on the application and fSampEn has not previously been applied to LS signals. This study aimed to perform an evaluation of the performance of the most relevant fSampEn parameters on LS signals, and to propose optimal fSampEn parameters for LSI estimation. Different combinations of fSampEn parameters were analyzed in LS signals recorded in a heterogeneous population of healthy subjects and chronic obstructive pulmonary disease patients during loaded breathing. The performance of fSampEn was assessed by means of its cross-covariance with flow signals, and optimal fSampEn parameters for LSI estimation were proposed.

Keywords: Large scale integration, Lung, Estimation, Entropy, Loading, Robustness, Diseases

Hortelão, Ana C., Carrascosa, Rafael, Murillo-Cremaes, Nerea, Patiño, Tania, Sánchez, Samuel, (2019). Targeting 3D bladder cancer spheroids with urease-powered nanomotors ACS Nano 13, (1), 429-439

Cancer is one of the main causes of death around the world, lacking efficient clinical treatments that generally present severe side effects. In recent years, various nanosystems have been explored to specifically target tumor tissues, enhancing the efficacy of cancer treatment and minimizing the side effects. In particular, bladder cancer is the ninth most common cancer worldwide and presents a high survival rate but serious recurrence levels, demanding an improvement in the existent therapies. Here, we present urease-powered nanomotors based on mesoporous silica nanoparticles that contain both polyethylene glycol and anti-FGFR3 antibody on their outer surface to target bladder cancer cells in the form of 3D spheroids. The autonomous motion is promoted by urea, which acts as fuel and is inherently present at high concentrations in the bladder. Antibody-modified nanomotors were able to swim in both simulated and real urine, showing a substrate-dependent enhanced diffusion. The internalization efficiency of the antibody-modified nanomotors into the spheroids in the presence of urea was significantly higher compared with antibody-modified passive particles or bare nanomotors. Furthermore, targeted nanomotors resulted in a higher suppression of spheroid proliferation compared with bare nanomotors, which could arise from the local ammonia production and the therapeutic effect of anti-FGFR3. These results hold significant potential for the development of improved targeted cancer therapy and diagnostics using biocompatible nanomotors.

Keywords: 3D cell culture, Bladder cancer, Enzymatic catalysis, Nanomachines, Nanomotors, Self-propulsion, Targeting

Kaurin, D., Arroyo, M., (2019). Surface tension controls the hydraulic fracture of adhesive interfaces bridged by molecular bonds Physical Review Letters 123, (22), 228102

Biological function requires cell-cell adhesions to tune their cohesiveness; for instance, during the opening of new fluid-filled cavities under hydraulic pressure. To understand the physical mechanisms supporting this adaptability, we develop a stochastic model for the hydraulic fracture of adhesive interfaces bridged by molecular bonds. We find that surface tension strongly enhances the stability of these interfaces by controlling flaw sensitivity, lifetime, and optimal architecture in terms of bond clustering. We also show that bond mobility embrittles adhesions and changes the mechanism of decohesion. Our study provides a mechanistic background to understand the biological regulation of cell-cell cohesion and fracture.

Keywords: Biomimetic & bio-inspired materials, Cell adhesion, Fracture, Self-healing

Roki, N., Tsinas, Z., Solomon, M., Bowers, J., Getts, R. C., Muro, S., (2019). Unprecedently high targeting specificity toward lung ICAM-1 using 3DNA nanocarriers Journal of Controlled Release 305, 41-49

DNA nanostructures hold great potential for drug delivery. However, their specific targeting is often compromised by recognition by scavenger receptors involved in clearance. In our previous study in cell culture, we showed targeting specificity of a 180 nm, 4-layer DNA-built nanocarrier called 3DNA coupled with antibodies against intercellular adhesion molecule-1 (ICAM-1), a glycoprotein overexpressed in the lungs in many diseases. Here, we examined the biodistribution of various 3DNA formulations in mice. A formulation consisted of 3DNA whose outer-layer arms were hybridized to secondary antibody-oligonucleotide conjugates. Anchoring IgG on this formulation reduced circulation and kidney accumulation vs. non-anchored IgG, while increasing liver and spleen clearance, as expected for a nanocarrier. Anchoring anti-ICAM changed the biodistribution of this antibody similarly, yet this formulation specifically accumulated in the lungs, the main ICAM-1 target. Since lung targeting was modest (2-fold specificity index over IgG formulation), we pursued a second preparation involving direct hybridization of primary antibody-oligonucleotide conjugates to 3DNA. This formulation had prolonged stability in serum and showed a dramatic increase in lung distribution: the specificity index was 424-fold above a matching IgG formulation, 144-fold more specific than observed for PLGA nanoparticles of similar size, polydispersity, ζ-potential and antibody valency, and its lung accumulation increased with the number of anti-ICAM molecules per particle. Immunohistochemistry showed that anti-ICAM and 3DNA components colocalized in the lungs, specifically associating with endothelial markers, without apparent histological changes. The degree of in vivo targeting for anti-ICAM/3DNA-nanocarriers is unprecedented, for which this platform technology holds great potential to develop future therapeutic applications.

Keywords: 3DNA, DNA nanostructure, Drug nanocarrier, Endothelial and lung targeting, ICAM-1, In vivo biodistribution

Manconi, M., Manca, M. L., Escribano-Ferrer, E., Coma-Cros, E. M., Biosca, A., Lantero, E., Fernàndez-Busquets, X., Fadda, A. M., Caddeo, C., (2019). Nanoformulation of curcumin-loaded eudragit-nutriosomes to counteract malaria infection by a dual strategy: Improving antioxidant intestinal activity and systemic efficacy International Journal of Pharmaceutics 556, 82-88

In this paper, nutriosomes (phospholipid vesicles associated with Nutriose® FM06) were modified to obtain new systems aimed at enhancing the efficacy of curcumin in counteracting malaria infection upon oral administration. Eudragit® L100, a pH-sensitive co-polymer, was added to these vesicles, thus obtaining eudragit-nutriosomes, to improve their in vivo performances. Liposomes without eudragit and nutriose were also prepared as a reference. Cryo-TEM images showed the formation of multicompartment vesicles, with mean diameter around 300 nm and highly negative zeta potential. Vesicles were stable in fluids mimicking the gastro-intestinal content due to the high phospholipid concentration and the presence of gastro-resistant eudragit and digestion-resistant nutriose. Eudragit-nutriosomes disclosed promising performances in vitro and in vivo: they maximized the ability of curcumin to counteract oxidative stress in intestinal cells (Caco-2), which presumably reinforced its systemic efficacy. Orally-administered curcumin-loaded eudragit-nutriosomes increased significantly the survival of malaria-infected mice relative to free curcumin-treated controls.

Keywords: Eudragit® L100, Nutriose® FM06, Nutriosomes, Curcumin, Oral administration, Malaria

Calvo, M., Le Rolle, V., Romero, D., Béhar, N., Gomis, P., Mabo, P., Hernández, A. I., (2019). Recursive model identification for the analysis of the autonomic response to exercise testing in Brugada syndrome Artificial Intelligence in Medicine 97, 98-104

This paper proposes the integration and analysis of a closed-loop model of the baroreflex and cardiovascular systems, focused on a time-varying estimation of the autonomic modulation of heart rate in Brugada syndrome (BS), during exercise and subsequent recovery. Patient-specific models of 44 BS patients at different levels of risk (symptomatic and asymptomatic) were identified through a recursive evolutionary algorithm. After parameter identification, a close match between experimental and simulated signals (mean error = 0.81%) was observed. The model-based estimation of vagal and sympathetic contributions were consistent with physiological knowledge, enabling to observe the expected autonomic changes induced by exercise testing. In particular, symptomatic patients presented a significantly higher parasympathetic activity during exercise, and an autonomic imbalance was observed in these patients at peak effort and during post-exercise recovery. A higher vagal modulation during exercise, as well as an increasing parasympathetic activity at peak effort and a decreasing vagal contribution during post-exercise recovery could be related with symptoms and, thus, with a worse prognosis in BS. This work proposes the first evaluation of the sympathetic and parasympathetic responses to exercise testing in patients suffering from BS, through the recursive identification of computational models; highlighting important trends of clinical relevance that provide new insights into the underlying autonomic mechanisms regulating the cardiovascular system in BS. The joint analysis of the extracted autonomic parameters and classic electrophysiological markers could improve BS risk stratification.

Keywords: Autonomic nervous system, Brugada syndrome, Computational model, Recursive identification

Ferrer, I., García, M. A., Carmona, M., Andrés-Benito, P., Torrejón-Escribano, B., Garcia-Esparcia, P., Del Rio, J. A., (2019). Involvement of oligodendrocytes in tau seeding and spreading in tauopathies Frontiers in Aging Neuroscience 11, 112

Introduction: Human tau seeding and spreading occur following intracerebral inoculation into different gray matter regions of brain homogenates obtained from tauopathies in transgenic mice expressing wild or mutant tau, and in wild-type (WT) mice. However, little is known about tau propagation following inoculation in the white matter. Objectives: The present study is geared to learning about the patterns of tau seeding and cells involved following unilateral inoculation in the corpus callosum of homogenates from sporadic Alzheimer's disease (AD), primary age-related tauopathy (PART: neuronal 4Rtau and 3Rtau), pure aging-related tau astrogliopathy (ARTAG: astroglial 4Rtau with thorn-shaped astrocytes TSAs), globular glial tauopathy (GGT: 4Rtau with neuronal tau and specific tau inclusions in astrocytes and oligodendrocytes, GAIs and GOIs, respectively), progressive supranuclear palsy (PSP: 4Rtau with neuronal inclusions, tufted astrocytes and coiled bodies), Pick's disease (PiD: 3Rtau with characteristic Pick bodies in neurons and tau containing fibrillar astrocytes), and frontotemporal lobar degeneration linked to P301L mutation (FTLD-P301L: 4Rtau familial tauopathy). Methods: Adult WT mice were inoculated unilaterally in the lateral corpus callosum with sarkosyl-insoluble fractions or with sarkosyl-soluble fractions from the mentioned tauopathies; mice were killed from 4 to 7 months after inoculation. Brains were fixed in paraformaldehyde, embedded in paraffin and processed for immunohistochemistry. Results: Tau seeding occurred in the ipsilateral corpus callosum and was also detected in the contralateral corpus callosum. Phospho-tau deposits were found in oligodendrocytes similar to coiled bodies and in threads. Moreover, tau deposits co-localized with active (phosphorylated) tau kinases p38 and ERK 1/2, suggesting active tau phosphorylation of murine tau. TSAs, GAIs, GOIs, tufted astrocytes, and tau-containing fibrillar astrocytes were not seen in any case. Tau deposits were often associated with slight myelin disruption and the presence of small PLP1-immunoreactive globules and dots in the ipsilateral corpus callosum 6 months after inoculation of sarkosyl-insoluble fractions from every tauopathy. Conclusions: Seeding and spreading of human tau in the corpus callosum of WT mice occurs in oligodendrocytes, thereby supporting the idea of a role of oligodendrogliopathy in tau seeding and spreading in the white matter in tauopathies. Slight differences in the predominance of threads or oligodendroglial deposits suggest disease differences in the capacity of tau seeding and spreading among tauopathies.

Keywords: AD, ARTAG, GGT, PiD, Seeding and spreading, Tau, Tauopathies

Lozano-García, M., Estrada-Petrocelli, L., Moxham, J., Rafferty, G. F., Torres, A., Jolley, C. J., Jané, R. , (2019). Noninvasive assessment of inspiratory muscle neuromechanical coupling during inspiratory threshold loading IEEE Access 7, 183634-183646

Diaphragm neuromechanical coupling (NMC), which reflects the efficiency of conversion of neural activation to transdiaphragmatic pressure (Pdi), is increasingly recognized to be a useful clinical index of diaphragm function and respiratory mechanics in neuromuscular weakness and cardiorespiratory disease. However, the current gold standard assessment of diaphragm NMC requires invasive measurements of Pdi and crural diaphragm electromyography (oesEMGdi), which complicates the measurement of diaphragm NMC in clinical practice. This is the first study to compare invasive measurements of diaphragm NMC (iNMC) using the relationship between Pdi and oesEMGdi, with noninvasive assessment of NMC (nNMC) using surface mechanomyography (sMMGlic) and electromyography (sEMGlic) of lower chest wall inspiratory muscles. Both invasive and noninvasive measurements were recorded in twelve healthy adult subjects during an inspiratory threshold loading protocol. A linear relationship between noninvasive sMMGlic and sEMGlic measurements was found, resulting in little change in nNMC with increasing inspiratory load. By contrast, a curvilinear relationship between invasive Pdi and oesEMGdi measurements was observed, such that there was a progressive increase in iNMC with increasing inspiratory threshold load. Progressive recruitment of lower ribcage muscles, serving to enhance the mechanical advantage of the diaphragm, may explain the more linear relationship between sMMGlic and sEMGlic (both representing lower intercostal plus costal diaphragm activity) than between Pdi and crural oesEMGdi. Noninvasive indices of NMC derived from sEMGlic and sMMGlic may prove to be useful indices of lower chest wall inspiratory muscle NMC, particularly in settings that do not have access to invasive measures of diaphragm function.

Keywords: Cardiovascular system, Diaphragms, Diseases, Electromyography, Medical signal processing, Neurophysiology, Patient monitoring, Pneumodynamics, Inspiratory muscle neuromechanical coupling, Diaphragm neuromechanical coupling, Neural activation, Transdiaphragmatic pressure, Diaphragm function, Respiratory mechanics, Diaphragm NMC, Invasive measurements, Crural diaphragm electromyography, iNMC, Noninvasive assessment, nNMC, Lower chest wall inspiratory muscles, Inspiratory threshold loading protocol, Noninvasive sMMGlic measurements, sEMGlic measurements, oesEMGdi measurements, Inspiratory threshold load, Lower ribcage muscles, Lower intercostal plus costal diaphragm activity, Crural oesEMGdi, Noninvasive indices, sEMGlic sMMGlic, Lower chest wall inspiratory muscle NMC, Surface mechanomyography, Electromyography, Inspiratory threshold loading, Mechanomyography, Neuromechanical coupling, Respiratory muscles

Herreros, Ivan, Miquel, Laia, Blithikioti, Chrysanthi, Nuño, Laura, Rubio Ballester, Belen, Grechuta, Klaudia, Gual, Antoni, Balcells-Oliveró, Mercè, Verschure, P., (2019). Motor adaptation impairment in chronic cannabis users assessed by a visuomotor rotation task Journal of Clinical Medicine 8, (7), 1049

Background—The cerebellum has been recently suggested as an important player in the addiction brain circuit. Cannabis is one of the most used drugs worldwide, and its long-term effects on the central nervous system are not fully understood. No valid clinical evaluations of cannabis impact on the brain are available today. The cerebellum is expected to be one of the brain structures that are highly affected by prolonged exposure to cannabis, due to its high density in endocannabinoid receptors. We aim to use a motor adaptation paradigm to indirectly assess cerebellar function in chronic cannabis users (CCUs). Methods—We used a visuomotor rotation (VMR) task that probes a putatively-cerebellar implicit motor adaptation process together with the learning and execution of an explicit aiming rule. We conducted a case-control study, recruiting 18 CCUs and 18 age-matched healthy controls. Our main measure was the angular aiming error. Results—Our results show that CCUs have impaired implicit motor adaptation, as they showed a smaller rate of adaptation compared with healthy controls (drift rate: 19.3 +/− 6.8° vs. 27.4 +/− 11.6°; t(26) = −2.1, p = 0.048, Cohen’s d = −0.8, 95% CI = (−1.7, −0.15)). Conclusions—We suggest that a visuomotor rotation task might be the first step towards developing a useful tool for the detection of alterations in implicit learning among cannabis users.

Keywords: Cerebellum, Cannabis, Implicit motor learning, Motor adaptation, Visuomotor rotation

Burgués, Javier, Hernández, Victor, Lilienthal, Achim J., Marco, Santiago, (2019). Smelling nano aerial vehicle for gas source localization and mapping Sensors 19, (3), 478

This paper describes the development and validation of the currently smallest aerial platform with olfaction capabilities. The developed Smelling Nano Aerial Vehicle (SNAV) is based on a lightweight commercial nano-quadcopter (27 g) equipped with a custom gas sensing board that can host up to two in situ metal oxide semiconductor (MOX) gas sensors. Due to its small form-factor, the SNAV is not a hazard for humans, enabling its use in public areas or inside buildings. It can autonomously carry out gas sensing missions of hazardous environments inaccessible to terrestrial robots and bigger drones, for example searching for victims and hazardous gas leaks inside pockets that form within the wreckage of collapsed buildings in the aftermath of an earthquake or explosion. The first contribution of this work is assessing the impact of the nano-propellers on the MOX sensor signals at different distances to a gas source. A second contribution is adapting the ‘bout’ detection algorithm, proposed by Schmuker et al. (2016) to extract specific features from the derivative of the MOX sensor response, for real-time operation. The third and main contribution is the experimental validation of the SNAV for gas source localization (GSL) and mapping in a large indoor environment (160 m2) with a gas source placed in challenging positions for the drone, for example hidden in the ceiling of the room or inside a power outlet box. Two GSL strategies are compared, one based on the instantaneous gas sensor response and the other one based on the bout frequency. From the measurements collected (in motion) along a predefined sweeping path we built (in less than 3 min) a 3D map of the gas distribution and identified the most likely source location. Using the bout frequency yielded on average a higher localization accuracy than using the instantaneous gas sensor response (1.38 m versus 2.05 m error), however accurate tuning of an additional parameter (the noise threshold) is required in the former case. The main conclusion of this paper is that a nano-drone has the potential to perform gas sensing tasks in complex environments.

Keywords: Robotics, Signal processing, Electronics, Gas source localization, Gas distribution mapping, Gas sensors, Drone, UAV, MOX sensor, Quadcopter

Sarlabous, L., Estrada, L., Cerezo-Hernández, A., Leest, Sietske V. D., Torres, A., Jané, R., Duiverman, M., Garde, Ainara, (2019). Electromyography-based respiratory onset detection in COPD patients on non-invasive mechanical ventilation Entropy 21, (3), 258

To optimize long-term nocturnal non-invasive ventilation in patients with chronic obstructive pulmonary disease, surface diaphragm electromyography (EMGdi) might be helpful to detect patient-ventilator asynchrony. However, visual analysis is labor-intensive and EMGdi is heavily corrupted by electrocardiographic (ECG) activity. Therefore, we developed an automatic method to detect inspiratory onset from EMGdi envelope using fixed sample entropy (fSE) and a dynamic threshold based on kernel density estimation (KDE). Moreover, we combined fSE with adaptive filtering techniques to reduce ECG interference and improve onset detection. The performance of EMGdi envelopes extracted by applying fSE and fSE with adaptive filtering was compared to the root mean square (RMS)-based envelope provided by the EMG acquisition device. Automatic onset detection accuracy, using these three envelopes, was evaluated through the root mean square error (RMSE) between the automatic and mean visual onsets (made by two observers). The fSE-based method provided lower RMSE, which was reduced from 298 ms to 264 ms when combined with adaptive filtering, compared to 301 ms provided by the RMS-based method. The RMSE was negatively correlated with the proposed EMGdi quality indices. Following further validation, fSE with KDE, combined with adaptive filtering when dealing with low quality EMGdi, indicates promise for detecting the neural onset of respiratory drive.

Keywords: Fixed sample entropy, Adaptive filtering, Root mean square, Diaphragm electromyography, Non-invasive mechanical ventilation, Chronic obstructive pulmonary disease

Campillo, N., Falcones, B., Otero, J., Colina, R., Gozal, D., Navajas, D., Farré, R., Almendros, I., (2019). Differential oxygenation in tumor microenvironment modulates macrophage and cancer cell crosstalk: Novel experimental settingand proof of concept Frontiers in Oncology 9, 43

Hypoxia is a common characteristic of many solid tumors that has been associated with tumor aggressiveness. Limited diffusion of oxygen generates a gradient of oxygen availability from the blood vessel to the interstitial space and may underlie the recruitment of macrophages fostering cancer progression. However, the available data based on the recruitment of circulating cells to the tumor microenvironment has been so far carried out by conventional co-culture systems which ignore the hypoxic gradient between the vessel to the tumor interstitium. Here, we have designed a novel easy-to-build cell culture device that enables evaluation of cellular cross-talk and cell migration while they are being simultaneously exposed to different oxygenation environments. As a proof-of-concept of the potential role of differential oxygenation among interacting cells we have evaluated the activation and recruitment of macrophages in response to hypoxic melanoma, breast, and kidney cancer cells. We found that hypoxic melanoma and breast cancer cells co-cultured with normoxic macrophages enhanced their directional migration. By contrast, hypoxic kidney cells were not able to increase their recruitment. We also identified well-described hypoxia-induced pathways which could contribute in the immune cell recruitment (VEGFA and PTGS2 genes). Moreover, melanoma and breast cancer increased their proliferation. However, oxygenation levels affected neither kidney cancer cell proliferation nor gene expression, which in turn resulted in no significant changes in macrophage migration and polarization. Therefore, the cell culture device presented here provides an excellent opportunity for researchers to reproduce the in vivo hypoxic gradients in solid tumors and to study their role in recruiting circulating cells to the tumor in specific types of cancer.

Keywords: Hypoxia gradient, Macrophage motility, Models of host-tumor interactions, Novel assay technology, Tumor progression

Ferrer, I., Zelaya, M. V., Aguiló García, M., Carmona, M., López-González, I., Andrés-Benito, P., Lidón, L., Gavín, R., Garcia-Esparcia, P., del Rio, J. A., (2019). Relevance of host tau in tau seeding and spreading in tauopathies Brain Pathology Early View

Human tau seeding and spreading occur following intracerebral inoculation of brain homogenates obtained from tauopathies in transgenic mice expressing natural or mutant tau, and in wild-type (WT) mice. The present study was geared to learning about the patterns of tau seeding, the cells involved and the characteristics of tau following intracerebral inoculation of homogenates from primary age-related tauopathy (PART: neuronal 4Rtau and 3Rtau), aging-related tau astrogliopathy (ARTAG: astrocytic 4Rtau) and globular glial tauopathy (GGT: 4Rtau with neuronal deposits and specific tau inclusions in astrocytes and oligodendrocytes). For this purpose, young and adult WT mice were inoculated unilaterally in the hippocampus or in the lateral corpus callosum with sarkosyl-insoluble fractions from PART, ARTAG and GGT cases, and were killed at variable periods of three to seven months. Brains were processed for immunohistochemistry in paraffin sections. Tau seeding occurred in the ipsilateral hippocampus and corpus callosum and spread to the septal nuclei, periventricular hypothalamus and contralateral corpus callosum, respectively. Tau deposits were mainly found in neurons, oligodendrocytes and threads; the deposits were diffuse or granular, composed of phosphorylated tau, tau with abnormal conformation and 3Rtau and 4Rtau independently of the type of tauopathy. Truncated tau at the aspartic acid 421 and ubiquitination were absent. Tau deposits had the characteristics of pre-tangles. A percentage of intracellular tau deposits co-localized with active (phosphorylated) tau kinases p38 and ERK 1/2. Present study shows that seeding and spreading of human tau into the brain of WT mice involves neurons and glial cells, mainly oligodendrocytes, thereby supporting the idea of a primary role of oligodendrogliopathy, together with neuronopathy, in the progression of tauopathies. In addition, it suggests that human tau inoculation modifies murine tau metabolism with the production and deposition of 3Rtau and 4Rtau, and by activation of specific tau kinases in affected cells.

Keywords: Aging-related tau astrogliopathy, Globular glial tauopathy, Primary age-related tauopathy, Seeding, Spreading, Tau, Tauopathies

Blanco-Almazán, D., Groenendaal, W., Catthoor, F., Jané, R., (2019). Analysis of time delay between bioimpedance and respiratory volume signals under inspiratory loaded breathing Engineering in Medicine and Biology Society (EMBC) 41st Annual International Conference of the IEEE , IEEE (Berlín, Germany) , 2365-2368

Bioimpedance is known for its linear relation with volume during normal breathing. For that reason, bioimpedance can be used as a noninvasive and comfortable technique for measuring respiration. The goal of this study is to analyze the temporal behavior of bioimpedance measured in four different electrode configurations during inspiratory loaded breathing. We measured four bioimpedance channels and airflow simultaneously in 10 healthy subjects while incremental inspiratory loads were imposed. Inspiratory loading threshold protocols are associated with breathing pattern changes and were used in respiratory mechanics studies. Consequently, this respiratory protocol allowed us to induce breathing pattern changes and evaluate the temporal relationship of bioimpedance with volume. We estimated the temporal delay between bioimpedance and volume respiratory cycles to evaluate the differences in their temporal behavior. The delays were computed as the lag which maximize the cross-correlation of the signals cycle by cycle. Six of the ten subjects showed delays in at least two different inspiratory loads. The delays were dependent on electrode configuration, hence the appearance of the delays between bioimpedance and volume were conditioned to the location and geometry of the electrode configuration. In conclusion, the delays between these signals could provide information about breathing pattern when breathing conditions change.

Keywords: Bioimpedance, Delays, Electrodes, Protocols, Loading, Electrocardiography, Atmospheric measurements

Ferrer-Lluis, I., Castillo-Escario, Y., Montserrat, J. M., Jané, R., (2019). Automatic event detector from smartphone accelerometry: Pilot mHealth study for obstructive sleep apnea monitoring at home Engineering in Medicine and Biology Society (EMBC) 41st Annual International Conference of the IEEE , IEEE (Berlín, Germany) , 4990-4993

Obstructive sleep apnea (OSA) is a common disorder with a low diagnosis ratio, leaving many patients undiagnosed and untreated. In the last decades, accelerometry has been found to be a feasible solution to obtain respiratory activity and a potential tool to monitor OSA. On the other hand, many smartphone-based systems have already been developed to propose solutions for OSA monitoring and treatment. The objective of this work was to develop an automatic event detector based on smartphone accelerometry and pulse oximetry, and to assess its ability to detect thoracic movements. It was validated with a commercial OSA monitoring system at home. Results of this preliminary pilot study showed that the proposed event detector for accelerometry signals is a feasible tool to detect abnormal respiratory events, such as apneas and hypopneas, and has potential to be included in smartphone-based systems for OSA assessment.

Keywords: Sleep apnea, Detectors, Pulse oximetry, Monitoring, Manuals, Band-pass filters, Pulse oximeter

Casanellas, Ignasi, Lagunas, Anna, Vida, Yolanda, Pérez-Inestrosa, Ezequiel, Andrades, José A., Becerra, José, Samitier, Josep, (2019). Matrix nanopatterning regulates mesenchymal differentiation through focal adhesion size and distribution according to cell fate Biomimetics Biomimetic Nanotechnology for Biomedical Applications (NanoBio&Med 2018) , MDPI (Barcelona, Spain) 4, (2), 43

Extracellular matrix remodeling plays a pivotal role during mesenchyme patterning into different lineages. Tension exerted from cell membrane receptors bound to extracellular matrix ligands is transmitted by the cytoskeleton to the cell nucleus inducing gene expression. Here, we used dendrimer-based arginine–glycine–aspartic acid (RGD) uneven nanopatterns, which allow the control of local surface adhesiveness at the nanoscale, to unveil the adhesive requirements of mesenchymal tenogenic and osteogenic commitments. Cell response was found to depend on the tension resulting from cell–substrate interactions, which affects nuclear morphology and is regulated by focal adhesion size and distribution.

Keywords: Arginine–glycine–aspartic acid (RGD), Nanopattern, Mesenchymal stem cells, Tenogenesis, Osteogenesis, Cell nuclei, Focal adhesions

Romero, D., Jané, R., (2019). Non-linear HRV analysis to quantify the effects of intermittent hypoxia using an OSA rat model Engineering in Medicine and Biology Society (EMBC) 41st Annual International Conference of the IEEE , IEEE (Berlín, Germany) , 4994-4997

In this paper, a non-linear HRV analysis was performed to assess fragmentation signatures observed in heartbeat time series after intermittent hypoxia (IH). Three markers quantifying short-term fragmentation levels, PIP, IALS and PSS, were evaluated on R-R interval series obtained in a rat model of recurrent apnea. Through airway obstructions, apnea episodes were periodically simulated in six anesthetized Sprague-Dawley rats. The number of apnea events per hour (AHI index) was varied during the first half of the experiment while apnea episodes lasted 15 s. For the second part, apnea episodes lasted 5, 10 or 15 s, but the AHI index was fixed. Recurrent apnea was repeated for 15-min time intervals in all cases, alternating with basal periods of the same duration. The fragmentation markers were evaluated in segments of 5 minutes, selected at the beginning and end of the experiment. The impact of the heart and breathing rates (HR and BR, respectively) on the parameter estimates was also investigated. The results obtained show a significant increase (from 5 to 10%, p <; 0.05) in fragmentation measures of heartbeat time series after IH, indicating a clear deterioration of the initial conditions. Moreover, there was a strong linear relationship (r > 0.9) between these markers and BR, as well as with the ratio given by HR/BR. Although fragmentation may be impacted by IH, we found that it is highly dependent on HR and BR values and thus, they should be considered during its calculation or used to normalize the fragmentation estimates.

Keywords: Rats, Time series analysis, Radio access technologies, Protocols, Heart beat

Martinez-Hernandez, Uriel, Vouloutsi, Vasiliki, Mura, Anna, Mangan, Michael, Asada, Minoru, Prescott, T. J., Verschure, P., (2019). Biomimetic and Biohybrid Systems 8th International Conference, Living Machines 2019, Nara, Japan, July 9–12, 2019, Proceedings , Springer, Cham (Lausanne, Switzerland) 11556, 1-384

This book constitutes the proceedings of the 8th International Conference on Biomimetic and Biohybrid Systems, Living Machines 2019, held in Nara, Japan, in July 2019. The 26 full and 16 short papers presented in this volume were carefully reviewed and selected from 45 submissions. They deal with research on novel life-like technologies inspired by the scientific investigation of biological systems, biomimetics, and research that seeks to interface biological and artificial systems to create biohybrid systems.

Keywords: Artificial intelligence, Biomimetics, Computer architecture, Human robot interaction, Human-Computer Interaction (HCI), Humanoid robot, Image processing, Learning algorithms, Mobile robots, Multipurpose robots, Neural networks, Quadruped robots, Reinforcement learning, Robot learning, Robotics, Robots, Sensor, Sensors, Swarm robotics, User interfaces

Hervera, A., De Virgiliis, F., Palmisano, I., Zhou, L., Tantardini, E., Kong, G., Hutson, T., Danzi, M. C., Perry, R. B. T., Santos, C. X. C., Kapustin, A. N., Fleck, R. A., Del Río, J. A., Carroll, T., Lemmon, V., Bixby, J. L., Shah, A. M., Fainzilber, M., Di Giovanni, S., (2018). Reactive oxygen species regulate axonal regeneration through the release of exosomal NADPH oxidase 2 complexes into injured axons Nature Cell Biology 20, (3), 307-319

Reactive oxygen species (ROS) contribute to tissue damage and remodelling mediated by the inflammatory response after injury. Here we show that ROS, which promote axonal dieback and degeneration after injury, are also required for axonal regeneration and functional recovery after spinal injury. We find that ROS production in the injured sciatic nerve and dorsal root ganglia requires CX3CR1-dependent recruitment of inflammatory cells. Next, exosomes containing functional NADPH oxidase 2 complexes are released from macrophages and incorporated into injured axons via endocytosis. Once in axonal endosomes, active NOX2 is retrogradely transported to the cell body through an importin-β1–dynein-dependent mechanism. Endosomal NOX2 oxidizes PTEN, which leads to its inactivation, thus stimulating PI3K–phosporylated (p-)Akt signalling and regenerative outgrowth. Challenging the view that ROS are exclusively involved in nerve degeneration, we propose a previously unrecognized role of ROS in mammalian axonal regeneration through a NOX2–PI3K–p-Akt signalling pathway.

Keywords: Adult neurogenesis, Endocytosis, Exocytosis, Monocytes and macrophages, Stress signalling

Del Río, J. A., Ferrer, Isidre, Gavín, R., (2018). Role of cellular prion protein in interneuronal amyloid transmission Progress in Neurobiology 165-167, 87-102

Several studies have indicated that certain misfolded amyloids composed of tau, β-amyloid or α-synuclein can be transferred from cell to cell, suggesting the contribution of mechanisms reminiscent of those by which infective prions spread through the brain. This process of a ‘prion-like’ spreading between cells is also relevant as a novel putative therapeutic target that could block the spreading of proteinaceous aggregates throughout the brain which may underlie the progressive nature of neurodegenerative diseases. The relevance of β-amyloid oligomers and cellular prion protein (PrPC) binding has been a focus of interest in Alzheimer’s disease (AD). At the molecular level, β-amyloid/PrPC interaction takes place in two differently charged clusters of PrPC. In addition to β-amyloid, participation of PrPC in α-synuclein binding and brain spreading also appears to be relevant in α-synucleopathies. This review summarizes current knowledge about PrPC as a putative receptor for amyloid proteins and the physiological consequences of these interactions..

Keywords: Cellular prion protein, Amyloid, Proteinaceous species, ‘prion-like’ spreading, Spreading, Neurodegeneration

Dix, Christina L., Matthews, Helen K., Uroz, Marina, McLaren, Susannah, Wolf, Lucie, Heatley, Nicholas, Win, Zaw, Almada, Pedro, Henriques, Ricardo, Boutros, Michael, Trepat, Xavier, Baum, Buzz, (2018). The role of mitotic cell-substrate adhesion re-modeling in animal cell division Developmental Cell 45, (1), 132-145

Animal cells undergo a dramatic series of shape changes as they divide, which depend on re-modeling of cell-substrate adhesions. Here, we show that while focal adhesion complexes are disassembled during mitotic rounding, integrins remain in place. These integrin-rich contacts connect mitotic cells to the underlying substrate throughout mitosis, guide polarized cell migration following mitotic exit, and are functionally important, since adherent cells undergo division failure when removed from the substrate. Further, the ability of cells to re-spread along pre-existing adhesive contacts is essential for division in cells compromised in their ability to construct a RhoGEF-dependent (Ect2) actomyosin ring. As a result, following Ect2 depletion, cells fail to divide on small adhesive islands but successfully divide on larger patterns, as the connection between daughter cells narrows and severs as they migrate away from one another. In this way, regulated re-modeling of cell-substrate adhesions during mitotic rounding aids division in animal cells.

Keywords: Division, Mitotic-rounding, Integrin-based adhesion, Cytokinesis

Sadowska, Joanna Maria, Guillem-Marti, Jordi, Espanol, Montserrat, Stähli, Christoph, Döbelin, Nicola, Ginebra, Maria-Pau, (2018). In vitro response of mesenchymal stem cells to biomimetic hydroxyapatite substrates: A new strategy to assess the effect of ion exchange Acta Biomaterialia 76, 319-332

Biomaterials can interact with cells directly, that is, by direct contact of the cells with the material surface, or indirectly, through soluble species that can be released to or uptaken from the surrounding fluids. However, it is difficult to characterise the relevance of this fluid-mediated interaction separately from the topography and composition of the substrate, because they are coupled variables. These fluid-mediated interactions are amplified in the case of highly reactive calcium phosphates (CaPs) such as biomimetic calcium deficient hydroxyapatite (CDHA), particularly in static in vitro cultures. The present work proposes a strategy to decouple the effect of ion exchange from topographical features by adjusting the volume ratio between the cell culture medium and biomaterial (VCM/VB). Increasing this ratio allowed mitigating the drastic ionic exchanges associated to the compositional changes experienced by the material exposed to the cell culture medium. This strategy was validated using rat mesenchymal stem cells (rMSCs) cultured on CDHA and beta-tricalcium phosphate (β-TCP) discs using different VCM/VB ratios. Whereas in the case of β-TCP the cell response was not affected by this ratio, a significant effect on cell adhesion and proliferation was found for the more reactive CDHA. The ionic exchange, produced by CDHA at low VCM/VB, altered cell adhesion due to the reduced number of focal adhesions, caused cell shrinkage and further rMCSs apoptosis. This was mitigated when using a high VCM/VB, which attenuated the changes of calcium and phosphate concentrations in the cell culture medium, resulting in rMSCs spreading and a viability over time. Moreover, rMSCs showed an earlier expression of osteogenic genes on CDHA compared to sintered β-TCP when extracellular calcium fluctuations were reduced. Statement of Significance: Fluid mediated interactions play a significant role in the bioactivity of calcium phosphates. Ionic exchange is amplified in the case of biomimetic hydroxyapatite, which makes the in vitro characterisation of cell-material interactions especially challenging. The present work proposes a novel and simple strategy to explore the mechanisms of interaction of biomimetic and sintered calcium phosphates with mesenchymal stem cells. The effects of topography and ion exchange are analysed separately by modifying the volume ratio between cell culture medium and biomaterial. High ionic fluctuations interfered in the maturation of focal adhesions, hampering cell adhesion and leading to increased apoptosis and reduced proliferation rate.

Keywords: Calcium phosphates, Mesenchymal stem cells, Intracellular calcium, Cell adhesion

González-García, C., Cantini, M., Ballester-Beltrán, J., Altankov, G., Salmerón-Sánchez, M., (2018). The strength of the protein-material interaction determines cell fate Acta Biomaterialia 77, 74-84

Extracellular matrix (ECM) proteins are key mediators of cell/material interactions. The surface density and conformation of these proteins adsorbed on the material surface influence cell adhesion and the cellular response. We have previously shown that subtle variations in surface chemistry lead to drastic changes in the conformation of adsorbed fibronectin (FN). On poly(ethyl acrylate) (PEA), FN unfolds and displays domains for cell adhesion and FN-FN interaction, whereas on poly(methyl acrylate) (PMA) – with only one methyl group less – FN remains globular as it is in solution. The effect of the strength of the protein/material interaction in cell response, and its relation to protein density and conformation, has received limited attention so far. In this work, we used FN-functionalized AFM cantilevers to evaluate, via force spectroscopy, the strength of interaction between fibronectin and the underlying polymer which controls FN conformation (PEA and PMA). We found that the strength of FN/PEA interaction is significantly higher than FN/PMA, which limits the mobility of FN layer on PEA, reduces the ability of cells to mechanically reorganize FN and then leads to enhanced proteolysis and degradation of the surrounding matrix with compromised cell viability. By contrast, both PEA and PMA support cell adhesion when FN density is increased and also in the presence of serum or other serum proteins, including vitronectin (VN) and bovine serum albumin (BSA), which provide a higher degree of mobility to the matrix. Statement of Significance: The identification of parameters influencing cell response is of paramount importance for the design of biomaterials that will act as synthetic scaffolds for cells to anchor, grow and, eventually, become specialised tissues. Cells interact with materials through an intermediate layer of proteins adsorbed on the material surface. It is known that the density and conformation of these proteins determine cell behaviour. Here we show that the strength of protein/material interactions, which has received very limited attention so far, is key to understand the cellular response to biomaterials. Very strong protein/material interactions reduce the ability of cells to mechanically reorganize proteins at the material interface which results in enhanced matrix degradation, leading ultimately to compromised cell viability.

Keywords: Fibronectin adsorption, Fibronectin remodeling, Protein mobility, Protein-material interaction strength

Urrea, L., Segura, Miriam, Masuda-Suzukake, M., Hervera, A., Pedraz, L., Aznar, J. M. G., Vila, M., Samitier, J., Torrents, E., Ferrer, Isidro, Gavín, R., Hagesawa, M., Del Río, J. A., (2018). Involvement of cellular prion protein in α-synuclein transport in neurons Molecular Neurobiology 55, (3), 1847-1860

The cellular prion protein, encoded by the gene Prnp, has been reported to be a receptor of β-amyloid. Their interaction is mandatory for neurotoxic effects of β-amyloid oligomers. In this study, we aimed to explore whether the cellular prion protein participates in the spreading of α-synuclein. Results demonstrate that Prnp expression is not mandatory for α-synuclein spreading. However, although the pathological spreading of α-synuclein can take place in the absence of Prnp, α-synuclein expanded faster in PrPC-overexpressing mice.

Keywords: Amyloid spreading, Microfluidic devices, Prnp, Synuclein

Sehgal, Poonam, Kong, Xinyu, Wu, Jun, Sunyer, Raimon, Trepat, Xavier, Leckband, Deborah, (2018). Epidermal growth factor receptor and integrins control force-dependent vinculin recruitment to E-cadherin junctions Journal of Cell Science 131, (6), jcs206656

This study reports novel findings that link E-cadherin (also known as CDH1)-mediated force-transduction signaling to vinculin targeting to intercellular junctions via epidermal growth factor receptor (EGFR) and integrins. These results build on previous findings that demonstrated that mechanically perturbed E-cadherin receptors activate phosphoinositide 3-kinase and downstream integrins in an EGFR-dependent manner. Results of this study show that this EGFR-mediated kinase cascade controls the force-dependent recruitment of vinculin to stressed E-cadherin complexes – a key early signature of cadherin-based mechanotransduction. Vinculin targeting requires its phosphorylation at tyrosine 822 by Abl family kinases (hereafter Abl), but the origin of force-dependent Abl activation had not been identified. We now present evidence that integrin activation, which is downstream of EGFR signaling, controls Abl activation, thus linking E-cadherin to Abl through a mechanosensitive signaling network. These findings place EGFR and integrins at the center of a positive-feedback loop, through which force-activated E-cadherin signals regulate vinculin recruitment to cadherin complexes in response to increased intercellular tension.This article has an associated First Person interview with the first author of the paper.

Keywords: Cadherin, Epidermal growth factor receptor, Force transduction, Magnetic twisting cytometry, Vinculin, Integrin

Crespo, Anna, Blanco-Cabra, N., Torrents, Eduard, (2018). Aerobic vitamin B12 biosynthesis is essential for pseudomonas aeruginosa class II ribonucleotide reductase activity during planktonic and biofilm growth Frontiers in Microbiology 9, (986), Article 986

P. aeruginosa is a major pathogenic bacterium in chronic infections and is a model organism for studying biofilms. P. aeruginosa is considered an aerobic bacterium, but in the presence of nitrate, it also grows in anaerobic conditions. Oxygen diffusion through the biofilm generates metabolic and genetic diversity in P. aeruginosa growth, such as in ribonucleotide reductase activity. These essential enzymes are necessary for DNA synthesis and repair. Oxygen availability determines the activity of the three-ribonucleotide reductase (RNR) classes. Class II and III RNRs are active in the absence of oxygen; however, class II RNRs, which are important in P. aeruginosa biofilm growth, require a vitamin B12 cofactor for their enzymatic activity. In this work, we elucidated the conditions in which class II RNRs are active due to vitamin B12 concentration constraints (biosynthesis or environmental availability). We demonstrated that increased vitamin B12 levels during aerobic, stationary and biofilm growth activate class II RNR activity. We also established that the cobN gene is essentially responsible for B12 biosynthesis under planktonic and biofilm growth. Our results unravel the mechanisms of dNTP synthesis by P. aeruginosa during biofilm growth, which appear to depend on the bacterial strain (laboratory-type or clinical isolate).

Keywords: Vitamin B12, Adenosylcobalamin, Ribonucleotide Reductases, Pseudomonas aeruginosa, NrdJ, Bacterial growth, Biofilm,Anaerobiosis

Martí Coma-Cros, Elisabet, Biosca, Arnau, Lantero, Elena, Manca, Maria, Caddeo, Carla, Gutiérrez, Lucía, Ramírez, Miriam, Borgheti-Cardoso, Livia, Manconi, Maria, Fernàndez-Busquets, Xavier, (2018). Antimalarial activity of orally administered curcumin incorporated in Eudragit®-containing liposomes International Journal of Molecular Sciences 19, (5), 1361

Curcumin is an antimalarial compound easy to obtain and inexpensive, having shown little toxicity across a diverse population. However, the clinical use of this interesting polyphenol has been hampered by its poor oral absorption, extremely low aqueous solubility and rapid metabolism. In this study, we have used the anionic copolymer Eudragit® S100 to assemble liposomes incorporating curcumin and containing either hyaluronan (Eudragit-hyaluronan liposomes) or the water-soluble dextrin Nutriose® FM06 (Eudragit-nutriosomes). Upon oral administration of the rehydrated freeze-dried nanosystems administered at 25/75 mg curcumin·kg−1·day−1, only Eudragit-nutriosomes improved the in vivo antimalarial activity of curcumin in a dose-dependent manner, by enhancing the survival of all Plasmodium yoelii-infected mice up to 11/11 days, as compared to 6/7 days upon administration of an equal dose of the free compound. On the other hand, animals treated with curcumin incorporated in Eudragit-hyaluronan liposomes did not live longer than the controls, a result consistent with the lower stability of this formulation after reconstitution. Polymer-lipid nanovesicles hold promise for their development into systems for the oral delivery of curcumin-based antimalarial therapies.

Keywords: Malaria, Curcumin, Nanomedicine, Oral administration, Lipid nanovesicles, Eudragit, Nutriose, Hyaluronan, Plasmodium yoelii

Silva, N., Riveros, A., Yutronic, N., Lang, E., Chornik, B., Guerrero, S., Samitier, J., Jara, P., Kogan, M. J., (2018). Photothermally controlled methotrexate release system using β-cyclodextrin and gold nanoparticles Nanomaterials 8, (12), 985

The inclusion compound (IC) of cyclodextrin (CD) containing the antitumor drug Methotrexate (MTX) as a guest molecule was obtained to increase the solubility of MTX and decrease its inherent toxic effects in nonspecific cells. The IC was conjugated with gold nanoparticles (AuNPs), obtained by a chemical method, creating a ternary intelligent delivery system for MTX molecules, based on the plasmonic properties of the AuNPs. Irradiation of the ternary system, with a laser wavelength tunable with the corresponding surface plasmon of AuNPs, causes local energy dissipation, producing the controlled release of the guest from CD cavities. Finally, cell viability was evaluated using MTS assays for β-CD/MTX and AuNPs + β-CD/MTX samples, with and without irradiation, against HeLa tumor cells. The irradiated sample of the ternary system AuNPs + β-CD/MTX produced a diminution in cell viability attributed to the photothermal release of MTX.

Keywords: Cyclodextrin, Delivery system, Gold nanoparticles, Inclusion compound, Irradiation, Laser, Methotrexate, Photothermal release

Mata, A., Gil, V., Pérez-Clausell, J., Dasilva, M., González-Calixto, M. C., Soriano, E., García-Verdugo, J. M., Sanchez-Vives, M. V., Del Río, J. A., (2018). New functions of Semaphorin 3E and its receptor PlexinD1 during developing and adult hippocampal formation Scientific Reports 8, (1), 1381

The development and maturation of cortical circuits relies on the coordinated actions of long and short range axonal guidance cues. In this regard, the class 3 semaphorins and their receptors have been seen to be involved in the development and maturation of the hippocampal connections. However, although the role of most of their family members have been described, very few data about the participation of Semaphorin 3E (Sema3E) and its receptor PlexinD1 during the development and maturation of the entorhino-hippocampal (EH) connection are available. In the present study, we focused on determining their roles both during development and in adulthood. We determined a relevant role for Sema3E/PlexinD1 in the layer-specific development of the EH connection. Indeed, mice lacking Sema3E/PlexinD1 signalling showed aberrant layering of entorhinal axons in the hippocampus during embryonic and perinatal stages. In addition, absence of Sema3E/PlexinD1 signalling results in further changes in postnatal and adult hippocampal formation, such as numerous misrouted ectopic mossy fibers. More relevantly, we describe how subgranular cells express PlexinD1 and how the absence of Sema3E induces a dysregulation of the proliferation of dentate gyrus progenitors leading to the presence of ectopic cells in the molecular layer. Lastly, Sema3E mutant mice displayed increased network excitability both in the dentate gyrus and the hippocampus proper.

Keywords: Adult neurogenesis, Axon and dendritic guidance

Taghadomi-Saberi, S., Garcia, S. M., Masoumi, A. A., Sadeghi, M., Marco, S., (2018). Classification of bitter orange essential oils according to fruit ripening stage by untargeted chemical profiling and machine learning Sensors 18, (6), 1922

The quality and composition of bitter orange essential oils (EOs) strongly depend on the ripening stage of the citrus fruit. The concentration of volatile compounds and consequently its organoleptic perception varies. While this can be detected by trained humans, we propose an objective approach for assessing the bitter orange from the volatile composition of their EO. The method is based on the combined use of headspace gas chromatography–mass spectrometry (HS-GC-MS) and artificial neural networks (ANN) for predictive modeling. Data obtained from the analysis of HS-GC-MS were preprocessed to select relevant peaks in the total ion chromatogram as input features for ANN. Results showed that key volatile compounds have enough predictive power to accurately classify the EO, according to their ripening stage for different applications. A sensitivity analysis detected the key compounds to identify the ripening stage. This study provides a novel strategy for the quality control of bitter orange EO without subjective methods.

Keywords: Bitter orange essential oil, Headspace gas chromatography–mass spectrometry, Artificial neural network, Foodomics, Chemometrics, Feature selection

Moulin-Frier, C., Fischer, T., Petit, M., Pointeau, G., Puigbo, J., Pattacini, U., Low, S. C., Camilleri, D., Nguyen, P., Hoffmann, M., Chang, H. J., Zambelli, M., Mealier, A., Damianou, A., Metta, G., Prescott, T. J., Demiris, Y., Dominey, P. F., Verschure, P. F. M. J., (2018). DAC-h3: A proactive robot cognitive architecture to acquire and express knowledge about the world and the self IEEE Transactions on Cognitive and Developmental Systems 10, (4), 1005-1022

This paper introduces a cognitive architecture for a humanoid robot to engage in a proactive, mixed-initiative exploration and manipulation of its environment, where the initiative can originate from both the human and the robot. The framework, based on a biologically-grounded theory of the brain and mind, integrates a reactive interaction engine, a number of state-of-the art perceptual and motor learning algorithms, as well as planning abilities and an autobiographical memory. The architecture as a whole drives the robot behavior to solve the symbol grounding problem, acquire language capabilities, execute goal-oriented behavior, and express a verbal narrative of its own experience in the world. We validate our approach in human-robot interaction experiments with the iCub humanoid robot, showing that the proposed cognitive architecture can be applied in real time within a realistic scenario and that it can be used with naive users.

Keywords: Autobiographical Memory., Biology, Cognition, Cognitive Robotics, Computer architecture, Distributed Adaptive Control, Grounding, Human-Robot Interaction, Humanoid robots, Robot sensing systems, Symbol Grounding

Truschzinski, M., Betella, A., Brunnett, G., Verschure, P., (2018). Emotional and cognitive influences in air traffic controller tasks: An investigation using a virtual environment? Applied Ergonomics 69, 1-9

Air traffic controllers are required to perform complex tasks which require attention and high precision. This study investigates how the difficulty of such tasks influences emotional states, cognitive workload and task performance. We use quantitative and qualitative measurements, including the recording of pupil dilation and changes in affect using questionnaires. Participants were required to perform a number of air traffic control tasks using the immersive human accessible Virtual Reality space in the "eXperience Induction Machine". Based on the data collected, we developed and validated a model which integrates personality, workload and affective theories. Our results indicate that the difficulty of an air traffic control task has a direct influence on cognitive workload as well as on the self-reported mood; whereas both mood and workload seem to change independently. In addition, we show that personality, in particular neuroticism, affects both mood and performance of the participants.

Keywords: Air traffic control, Mood, Personality, Virtual reality, Workload

Miquel, Joan, Santana, F., Palau, E., Vinagre, M., Langohr, K., Casals, A., Torrens, C., (2018). Retaining or excising the supraspinatus tendon in complex proximal humeral fractures treated with reverse prosthesis: a biomechanical analysis in two different designs Archives of Orthopaedic and Trauma Surgery 138, (11), 1533-1539

We aimed to biomechanically evaluate the effect of the supraspinatus tendon on tuberosity stability using two different reverse shoulder arthroplasty (RSA) models for complex proximal humeral fractures (PHFs).

Keywords: Tuberosity reconstruction, Reverse shoulder arthroplasty, Supraspinatus, Cadaveric study, Rotator cuff excision, Complex proximal humeral fractures

Casanellas, Ignasi, Lagunas, Anna, Tsintzou, Iro, Vida, Yolanda, Collado, Daniel, Pérez-Inestrosa, Ezequiel, Rodríguez-Pereira, Cristina, Magalhaes, Joana, Gorostiza, Pau, Andrades, José A., Becerra, José, Samitier, Josep, (2018). Dendrimer-based uneven nanopatterns to locally control surface adhesiveness: A method to direct chondrogenic differentiation Journal of Visualized Experiments Bioengineering, (131), e56347

Cellular adhesion and differentiation is conditioned by the nanoscale disposition of the extracellular matrix (ECM) components, with local concentrations having a major effect. Here we present a method to obtain large-scale uneven nanopatterns of arginine-glycine-aspartic acid (RGD)-functionalized dendrimers that permit the nanoscale control of local RGD surface density. Nanopatterns are formed by surface adsorption of dendrimers from solutions at different initial concentrations and are characterized by water contact angle (CA), X-ray photoelectron spectroscopy (XPS), and scanning probe microscopy techniques such as scanning tunneling microscopy (STM) and atomic force microscopy (AFM). The local surface density of RGD is measured using AFM images by means of probability contour maps of minimum interparticle distances and then correlated with cell adhesion response and differentiation. The nanopatterning method presented here is a simple procedure that can be scaled up in a straightforward manner to large surface areas. It is thus fully compatible with cell culture protocols and can be applied to other ligands that exert concentration-dependent effects on cells.

Keywords: Bioengineering, Dendrimer, Nanopattern, Arginine-Glycine-Aspartic Acid (RGD), Atomic Force Microscopy (AFM), Cell Adhesion, Mesenchymal Stem Cells (Mscs), Chondrogenesis

Verschure, P., (2018). A chronology of Distributed Adaptive Control Living machines: A handbook of research in biomimetics and biohybrid systems (ed. Prescott, T. J., Lepora, Nathan, Verschure, P.), Oxford Scholarship (Oxford, UK) , 346-360

This chapter presents the Distributed Adaptive Control (DAC) theory of the mind and brain of living machines. DAC provides an explanatory framework for biological brains and an integration framework for synthetic ones. DAC builds on several themes presented in the handbook: it integrates different perspectives on mind and brain, exemplifies the synthetic method in understanding living machines, answers well-defined constraints faced by living machines, and provides a route for the convergent validation of anatomy, physiology, and behavior in our explanation of biological living machines. DAC addresses the fundamental question of how a living machine can obtain, retain, and express valid knowledge of its world. We look at the core components of DAC, specific benchmarks derived from the engagement with the physical and the social world (the H4W and the H5W problems) in foraging and human–robot interaction tasks. Lastly we address how DAC targets the UTEM benchmark and the relation with contemporary developments in AI.

Keywords: Distributed Adaptive Control, Problem of priors, Symbol grounding problem, Convergent validation, Foraging, brain, Architecture, system

Lepora, Nathan, Verschure, P., Prescott, T. J., (2018). A roadmap for Living Machines research Living machines: A handbook of research in biomimetics and biohybrid systems (ed. Prescott, T. J., Lepora, Nathan, Verschure, P.), Oxford Scholarship (Oxford, UK) , 26-50

This roadmap identifies current trends in biomimetic and biohybrid systems together with their implications for future research and innovation. Important questions include the scale at which these systems are defined, the types of biological systems addressed, the kind of principles sought, the differences between biologically based and biologically inspired approaches, the role in the understanding of living systems, relevant application domains, common benchmarks, the relation to other fields, and developments on the horizon. We interviewed and collated answers from experts who have been involved a series of events organized by the Convergent Science Network. These answers were then collated into themes of research. Overall, we see a field rapidly expanding in influence and impact. As such, this report will provide information to researchers and scientific policy makers on contemporary biomimetics and its future, together with pointers to further reading on relevant topics within this handbook.

Keywords: Biomimetics, Biohybrid, Bio-inspiration, Biologically inspired, Roadmap, Living machines, policy

Verschure, P., Prescott, T. J., (2018). A living machines approach to the sciences of mind and brain Living Machines: A Handbook of Research in Biomimetic and Biohybrid Systems (ed. Prescott, T. J., Lepora, Nathan, Verschure, P.), Oxford Scholarship (Oxford, UK) , 15-25

How do the sciences of mind and brain—neuroscience, psychology, cognitive science, and artificial intelligence (AI)—stand in relation to each other in the 21st century? This chapter proposes that despite our knowledge expanding at ever-accelerating rates, our understanding of the relationship between mind and brain is, in some important sense, becoming less and less. An increasing explanatory gap can only be bridged by a multi-tiered and integrated theoretical framework that recognizes the value of developing explanations at different levels, combining these into cross-level integrated theories, and directly contributing to new technologies that improve the human condition. Development of technologies that instantiate principles gleaned from the study of the mind and brain, or biomimetic technologies, is a key part of the validation process for scientific theories of mind and brain. We call this strategy for the integration of science and engineering a Living Machines approach. Following this path can lead not only to better science, and useful engineering, but also a richer view of human experience and of relationships between science, engineering, and art.

Keywords: Convergent validation, Multi-tiered theories, Paradigms in cognitive science, Philosophy of science, Physical models, Reductionism

Hristova-Panusheva, K., Keremidarska-Markova, M., Altankov, G., Krasteva, N., (2017). Age-related changes in adhesive phenotype of bone marrow-derived mesenchymal stem cells on extracellular matrix proteins Journal of New Results in Science , 6, (1), 11-19

Mesenchymal stem cells (MSCs) are a promising cell source for cell-based therapies because of their self-renewal and multi-lineage differentiation potential. Unlike embryonic stem cells adult stem cells are subject of aging processes and the concomitant decline in their function. Age-related changes in MSCs have to be well understood in order to develop clinical techniques and therapeutics based on these cells. In this work we have studied the effect of aging on adhesive behaviour of bone marrow-derived MSC and MG- 63 osteoblastic cells onto three extracellular matrix proteins: fibronectin (FN), vitronectin (VN) and collagen I (Coll I). The results revealed substantial differences in adhesive behaviour of both cell types during 21 days in culture. Bone-marrow derived MSCs decreased significantly their adhesive affinity to all studied proteins after 7th day in culture with further incubation. In contrast, MG-63 cells, demonstrated a stable cell adhesive phenotype with high affinity to FN and Coll I and low affinity to vitronectin over the whole culture period. These data suggest that adhesive behaviour of MSCs to matrix proteins is affected by aging processes unlike MG-63 cells and the age-related changes have to be considered when expanding adult stem cells for clinical applications.

Keywords: Cell morphology, Cell attachment and spreading, Fibronectin, Vitronectin, Collagen I

Neri, L., Lasa, M., Elosegui-Artola, A., D'Avola, D., Carte, B., Gazquez, C., Alve, S., Roca-Cusachs, P., Iñarrairaegui, M., Herrero, J., Prieto, J., Sangro, B., Aldabe, R., (2017). NatB-mediated protein N-α-terminal acetylation is a potential therapeutic target in hepatocellular carcinoma Oncotarget 8, (25), 40967-40981

The identification of new targets for systemic therapy of hepatocellular carcinoma (HCC) is an urgent medical need. Recently, we showed that hNatB catalyzes the N-α-terminal acetylation of 15% of the human proteome and that this action is necessary for proper actin cytoskeleton structure and function. In tumors, cytoskeletal changes influence motility, invasion, survival, cell growth and tumor progression, making the cytoskeleton a very attractive antitumor target. Here, we show that hNatB subunits are upregulated in in over 59% HCC tumors compared to non-tumor tissue and that this upregulation is associated with microscopic vascular invasion. We found that hNatB silencing blocks proliferation and tumor formation in HCC cell lines in association with hampered DNA synthesis and impaired progression through the S and the G2/M phases. Growth inhibition is mediated by the degradation of two hNatB substrates, tropomyosin and CDK2, which occurs when these proteins lack N-α-terminal acetylation. In addition, hNatB inhibition disrupts the actin cytoskeleton, focal adhesions and tight/adherens junctions, abrogating two proliferative signaling pathways, Hippo/YAP and ERK1/2. Therefore, inhibition of NatB activity represents an interesting new approach to treating HCC by blocking cell proliferation and disrupting actin cytoskeleton function.

Keywords: CDK2, Cell cycle arrest, Cell-cell junctions, Focal adhesions, Tropomyosin

Parmar, J., Villa, K., Vilela, D., Sánchez, S., (2017). Platinum-free cobalt ferrite based micromotors for antibiotic removal Applied Materials Today 9, 605-611

Self-propelled micromotors have previously shown to enhance pollutant removal compared to non-motile nano-micro particles. However, these systems are expensive, difficult to scale-up and require surfactant for efficient work. Efficient and inexpensive micromotors are desirable for their practical applications in water treatment technologies. We describe cobalt-ferrite based micromotors (CFO micromotors) fabricated by a facile and scalable synthesis, that produce hydroxyl radicals via Fenton-like reaction and take advantage of oxygen gas generated during this reaction for self-propulsion. Once the reaction is complete, the CFO micromotors can be easily separated and collected due to their magnetic nature. The CFO micromotors are demonstrated for highly efficient advanced oxidative removal of tetracycline antibiotic from the water. Furthermore, the effects of different concentrations of micromotors and hydrogen peroxide on the antibiotic degradation were studied, as well as the generation of the highly reactive hydroxyl radicals responsible for the oxidation reaction.

Keywords: Degradation, Fenton reaction, Microbots, Nanomotors, Self-propelled Micromotors, Water treatment

Ojosnegros', Samuel, Cutrale, Francesco, Rodríguez, Daniel, Otterstrom, Jason J., Chiu, Chi Li, Hortigüela, Verónica, Tarantino, Carolina, Seriola', Anna, Mieruszynski, Stephen, Martínez, Elena, Lakadamyali, Melike, Raya, Angel, Fraser, Scott E., (2017). Eph-ephrin signaling modulated by polymerization and condensation of receptors Proceedings of the National Academy of Sciences of the United States of America 114, (50), 13188-13193

Eph receptor signaling plays key roles in vertebrate tissue boundary formation, axonal pathfinding, and stem cell regeneration by steering cells to positions defined by its ligand ephrin. Some of the key events in Eph-ephrin signaling are understood: ephrin binding triggers the clustering of the Eph receptor, fostering transphosphorylation and signal transduction into the cell. However, a quantitative and mechanistic understanding of how the signal is processed by the recipient cell into precise and proportional responses is largely lacking. Studying Eph activation kinetics requires spatiotemporal data on the number and distribution of receptor oligomers, which is beyond the quantitative power offered by prevalent imaging methods. Here we describe an enhanced fluorescence fluctuation imaging analysis, which employs statistical resampling to measure the Eph receptor aggregation distribution within each pixel of an image. By performing this analysis over time courses extending tens of minutes, the information-rich 4D space (x, y, oligomerization, time) results were coupled to straightforward biophysical models of protein aggregation. This analysis reveals that Eph clustering can be explained by the combined contribution of polymerization of receptors into clusters, followed by their condensation into far larger aggregates. The modeling reveals that these two competing oligomerization mechanisms play distinct roles: polymerization mediates the activation of the receptor by assembling monomers into 6- to 8-mer oligomers; condensation of the preassembled oligomers into large clusters containing hundreds of monomers dampens the signaling. We propose that the polymerization–condensation dynamics creates mechanistic explanation for how cells properly respond to variable ligand concentrations and gradients.

Keywords: Eph, Ephrin, Receptor tyrosine kinase, Gradients, Cell communication

Hoyos-Nogués, M., Velasco, F., Ginebra, M. P., Manero, J. M., Gil, F. J., Mas-Moruno, C., (2017). Regenerating bone via multifunctional coatings: The blending of cell integration and bacterial inhibition properties on the surface of biomaterials ACS Applied Materials & Interfaces 9, (26), 21618-21630

In dentistry and orthopedics, it is well accepted that implant fixation is a major goal. However, an emerging concern is bacterial infection. Infection of metallic implants can be catastrophic and significantly reduce patient quality of life. Accordingly, in this work, we focus on multifunctional coatings to simultaneously address and mitigate both these problems. We have developed a tailor-made peptide-based chemical platform that integrates the well-known RGD cell adhesive sequence and the lactoferrin-derived LF1-11 antimicrobial peptide. The platform was covalently grafted on titanium via silanization and the functionalization process characterized by contact angle, XPS, and QCM-D. The presence of the platform statistically improved the adhesion, proliferation and mineralization of osteoblast-like cells compared to control surfaces. At the same time, colonization by representative bacterial strains was significantly reduced on the surfaces. Furthermore, the biological potency of the multifunctional platform was verified in a co-culture in vitro model. Our findings demonstrate that this multifunctional approach can be useful to functionalize biomaterials to both improve cell integration and reduce the risk of bacterial infection.

Keywords: Antimicrobial peptides, Cell adhesive peptides, Multifunctionality, Osseointegration, Surface functionalization

Matalonga, J., Glaria, E., Bresque, M., Escande, C., Carbó, J. M., Kiefer, K., Vicente, R., León, T. E., Beceiro, S., Pascual-García, M., Serret, J., Sanjurjo, L., Morón-Ros, S., Riera, A., Paytubi, S., Juarez, A., Sotillo, F., Lindbom, L., Caelles, C., Sarrias, M. R., Sancho, J., Castrillo, A., Chini, E. N., Valledor, A. F., (2017). The nuclear receptor LXR limits bacterial infection of host macrophages through a mechanism that impacts cellular NAD metabolism Cell Reports 18, (5), 1241-1255

Macrophages exert potent effector functions against invading microorganisms but constitute, paradoxically, a preferential niche for many bacterial strains to replicate. Using a model of infection by Salmonella Typhimurium, we have identified a molecular mechanism regulated by the nuclear receptor LXR that limits infection of host macrophages through transcriptional activation of the multifunctional enzyme CD38. LXR agonists reduced the intracellular levels of NAD+ in a CD38-dependent manner, counteracting pathogen-induced changes in macrophage morphology and the distribution of the F-actin cytoskeleton and reducing the capability of non-opsonized Salmonella to infect macrophages. Remarkably, pharmacological treatment with an LXR agonist ameliorated clinical signs associated with Salmonella infection in vivo, and these effects were dependent on CD38 expression in bone-marrow-derived cells. Altogether, this work reveals an unappreciated role for CD38 in bacterial-host cell interaction that can be pharmacologically exploited by activation of the LXR pathway.

Keywords: Bacterial infection, CD38, Cytoskeleton, LXR, Macrophage, NAD, Nuclear receptor

Diez-Escudero, A., Espanol, M., Beats, S., Ginebra, M. P., (2017). In vitro degradation of calcium phosphates: Effect of multiscale porosity, textural properties and composition Acta Biomaterialia 60, 81-92

The capacity of calcium phosphates to be replaced by bone is tightly linked to their resorbability. However, the relative importance of some textural parameters on their degradation behavior is still unclear. The present study aims to quantify the effect of composition, specific surface area (SSA), and porosity at various length scales (nano-, micro- and macroporosity) on the in vitro degradation of different calcium phosphates. Degradation studies were performed in an acidic medium to mimic the osteoclastic environment. Small degradations were found in samples with interconnected nano- and micropores with sizes below 3 µm although they were highly porous (35–65%), with maximum weight loss of 8 wt%. Biomimetic calcium deficient hydroxyapatite, with high SSA and low crystallinity, presented the highest degradation rates exceeding even the more soluble β-TCP. A dependence of degradation on SSA was indisputable when porosity and pore sizes were increased. The introduction of additional macroporosity with pore interconnections above 20 µm significantly impacted degradation, more markedly in the substrates with high SSA (>15 m2/g), whereas in sintered substrates with low SSA (<1 m2/g) it resulted just in a linear increase of degradation. Up to 30 % of degradation was registered in biomimetic substrates, compared to 15 % in β-TCP or 8 % in sintered hydroxyapatite. The incorporation of carbonate in calcium deficient hydroxyapatite did not increase its degradation rate. Overall, the study highlights the importance of textural properties, which can modulate or even outweigh the effect of other features such as the solubility of the compounds. Statement of Significance The physicochemical features of calcium phosphates are crucial to tune biological events like resorption during bone remodeling. Understanding in vitro resorption can help to predict the in vivo behavior. Besides chemical composition, other parameters such as porosity and specific surface area have a strong influence on resorption. The complexity of isolating the contribution of each parameter lies in the close interrelation between them. In this work, a multiscale study was proposed to discern the extent to which each parameter influences degradation in a variety of calcium phosphates, using an acidic medium to resemble the osteoclastic environment. The results emphasize the importance of textural properties, which can modulate or even outweigh the effect of the intrinsic solubility of the compounds.

Keywords: Calcium phosphates, Degradation, Porosity, Textural properties

Bianchi, M. V., Awaja, F., Altankov, G., (2017). Dynamic adhesive environment alters the differentiation potential of young and ageing mesenchymal stem cells Materials Science and Engineering: C 78, 467-474

Engineering dynamic stem cell niche-like environment offers opportunity to obtain better control of the fate of stem cells. We identified, for the first time, that periodic changes in the adhesive environment of human adipose derived mesenchymal stem cells (ADSCs) alters dramatically their asymmetric division but not their ability for symmetric renewal. Hereby, we used smart thermo-responsive polymer (PNIPAM) to create a dynamic adhesive environment for ADSCs by applying periodic temperature cycles to perturb adsorbed adhesive proteins to substratum interaction. Cumulative population doubling time (CPDT) curves showed insignificant decline in the symmetric cell growth studied for up to 13th passages accompanied with small changes in the overall cell morphology and moderately declined fibronectin (FN) matrix deposition probably as a functional consequence of ADSCs ageing. However, a substantial alteration in the differentiation potential of ADSCs from both early and late passages (3rd and 14th, respectively) was found when the cells were switched to osteogenic differentiation conditions. This behavior was evidenced by the significantly altered alkaline phosphatase activity and Ca deposition (Alizarin red) assayed at 3, 14 and 21 day in comparison to the control samples of regular TC polystyrene processed under same temperature settings.

Keywords: Cell ageing, Dynamic adhesive environment, Extracellular matrix, Mesenchymal stem cells, PNIPAM, Stem cell niche, Symmetric and asymmetric cell growth, Thermo-cycling, Thermo-responsive polymer

Castellanos, M. I., Mas-Moruno, C., Grau, A., Serra-Picamal, X., Trepat, X., Albericio, F., Joner, M., Gil, F. J., Ginebra, M. P., Manero, J. M., Pegueroles, M., (2017). Functionalization of CoCr surfaces with cell adhesive peptides to promote HUVECs adhesion and proliferation Applied Surface Science , 393, 82-92

Biomimetic surface modification with peptides that have specific cell-binding moieties is a promising approach to improve endothelialization of metal-based stents. In this study, we functionalized CoCr surfaces with RGDS, REDV, YIGSR peptides and their combinations to promote endothelial cells (ECs) adhesion and proliferation. An extensive characterization of the functionalized surfaces was performed by XPS analysis, surface charge and quartz crystal microbalance with dissipation monitoring (QCM-D), which demonstrated the successful immobilization of the peptides to the surface. Cell studies demonstrated that the covalent functionalization of CoCr surfaces with an equimolar combination of RGDS and YIGSR represents the most powerful strategy to enhance the early stages of ECs adhesion and proliferation, indicating a positive synergistic effect between the two peptide motifs. Although these peptide sequences slightly increased smooth muscle cells (SMCs) adhesion, these values were ten times lower than those observed for ECs. The combination of RGDS with the REDV sequence did not show synergistic effects in promoting the adhesion or proliferation of ECs. The strategy presented in this study holds great potential to overcome clinical limitations of current metal stents by enhancing their capacity to support surface endothelialization.

Keywords: Cell adhesive peptides, CoCr alloy, Endothelialization, HUVEC proliferation, SMCs adhesion, Surface functionalization

Sachot, N., Roguska, A., Planell, J. P., Lewandowska, M., Engel, E., Castaño, O., (2017). Fast-degrading PLA/ORMOGLASS fibrous composite scaffold leads to a calcium-rich angiogenic environment International Journal of Nanomedicine 12, 4901-4919

The success of scaffold implantation in acellular tissue engineering approaches relies on the ability of the material to interact properly with the biological environment. This behavior mainly depends on the design of the graft surface and, more precisely, on its capacity to biodegrade in a well-defined manner (nature of ions released, surface-to-volume ratio, dissolution profile of this release, rate of material resorption, and preservation of mechanical properties). The assessment of the biological behavior of temporary templates is therefore very important in tissue engineering, especially for composites, which usually exhibit complicated degradation behavior. Here, blended polylactic acid (PLA) calcium phosphate ORMOGLASS (organically modified glass) nanofibrous mats have been incubated up to 4 weeks in physiological simulated conditions, and their morphological, topographical, and chemical changes have been investigated. The results showed that a significant loss of inorganic phase occurred at the beginning of the immersion and the ORMOGLASS maintained a stable composition afterward throughout the degradation period. As a whole, the nanostructured scaffolds underwent fast and heterogeneous degradation. This study reveals that an angiogenic calcium-rich environment can be achieved through fast-degrading ORMOGLASS/PLA blended fibers, which seems to be an excellent alternative for guided bone regeneration.

Keywords: Angiogenesis, Calcium release, Electrospinning, Fast degradation, Nanofibers, ORMOGLASSES

Castellanos, M. I., Guillem-Marti, J., Mas-Moruno, C., Díaz-Ricart, M., Escolar, G., Ginebra, M. P., Gil, F. J., Pegueroles, M., Manero, J. M., (2017). Cell adhesive peptides functionalized on CoCr alloy stimulate endothelialization and prevent thrombogenesis and restenosis Journal of Biomedical Materials Research - Part A , 105, (4), 973-983

Immobilization of bioactive peptide sequences on CoCr surfaces is an effective route to improve endothelialization, which is of great interest for cardiovascular stents. In this work, we explored the effect of physical and covalent immoblization of RGDS, YIGSR and their equimolar combination peptides on endothelial cells (EC) and smooth muscle cell (SMC) adhesion and on thrombogenicity. We extensively investigated using RT-qPCR, the expression by ECs cultured on functionalised CoCr surfaces of different genes. Genes relevant for adhesion (ICAM-1 and VCAM-1), vascularization (VEGFA, VEGFR-1 and VEGFR-2) and anti-thrombogenicity (tPA and eNOS) were over-expressed in the ECs grown to covalently functionalized CoCr surfaces compared to physisorbed and control surfaces. Pro-thrombogenic genes expression (PAI-1 and vWF) decreased over time. Cell co-cultures of ECs/SMCs found that functionalization increased the amount of adhered ECs onto modified surfaces compared to plain CoCr, independently of the used peptide and the strategy of immobilization. SMCs adhered less compared to ECs in all surfaces. All studied peptides showed a lower platelet cell adhesion compared to TCPS. Covalent functionalization of CoCr surfaces with an equimolar combination of RGDS and YIGSR represented prevailing strategy to enhance the early stages of ECs adhesion and proliferation, while preventing SMCs and platelet adhesion.

Keywords: Cell coculture, CoCr alloy, Functionalization, Gene expression, Platelet adhesion

Garcia-Castellote, D., Torres, A., Estrada, L., Sarlabous, L., Jane, R., (2017). Evaluation of indirect measures of neural inspiratory time from invasive and noninvasive recordings of respiratory activity Engineering in Medicine and Biology Society (EMBC) 39th Annual International Conference of the IEEE , IEEE (Seogwipo, South Korea) , 341-344

Measuring diaphragmatic electromyography (EMGdi) provides an indirect quantification of neural respiratory drive and allows the delimitation of diaphragm neural activation and deactivation during inspiration. EMGdi recordings have been incorporated in novel modes of assisted mechanical ventilation, such as neurally adjusted ventilatory assist (NAVA), to trigger and cycle-off the ventilator. The EMGdi signal improves the assistance delivered by more conventional ventilatory modes, in which the ventilator is synchronized with the patient employing a pneumatic triggering. In this work, we evaluate the time delay between the onset and offset of inspiratory activity estimated from EMGdi and three respiratory mechanical signals: the respiratory flow (FL), the transdiaphragmatic pressure (Pdi) and the diaphragm length (Ldi) signals. To this purpose, these signals were acquired in three mongrel dogs surgically instrumented under general anesthesia. Onsets and offsets were estimated manually and by automatic algorithms on these signals. The highest delays were obtained between EMGdi and FL (100 ms) while the lowest delays were obtained between EMGdi and Pdi (8 ms). Moreover, differences between manual and automatic estimations showed a mean absolute error lower than 45 ms. In conclusion, our study points out that both EMGdi and Pdi signals detect the onset and offset of inspiratory activity earlier than the FL signal, and would therefore be better for the improvement of patient-ventilator synchrony.

Keywords: Estimation, Ventilation, Anesthesia, Dogs, Manuals, Power harmonic filters

Quadri, M., Matera, C., Silnovi, Pismataro, M. C., Horenstein, N. A., Stokes, C., Papke, R. L., Dallanoce, C., (2017). Identification of α7 nicotinic acetylcholine receptor silent agonists based on the spirocyclic quinuclidine-Δ2-isoxazoline scaffold: Synthesis and electrophysiological evaluation ChemMedChem XXIV National Meeting in Medicinal Chemistry (NMMC 2016) , Wiley Online Library (Perugia, Spain) 12, (16), 1335-1348

Compound 11 (3-(benzyloxy)-1′-methyl-1′-azonia-4H-1′-azaspiro[isoxazole-5,3′-bicyclo[2.2.2]octane] iodide) was selected from a previous set of nicotinic ligands as a suitable model compound for the design of new silent agonists of α7 nicotinic acetylcholine receptors (nAChRs). Silent agonists evoke little or no channel activation but can induce the α7 desensitized Ds state, which is sensitive to a type II positive allosteric modulator, such as PNU-120596. Introduction of meta substituents into the benzyloxy moiety of 11 led to two sets of tertiary amines and quaternary ammonium salts based on the spirocyclic quinuclidinyl-Δ2-isoxazoline scaffold. Electrophysiological assays performed on Xenopus laevis oocytes expressing human α7 nAChRs highlighted four compounds that are endowed with a significant silent-agonism profile. Structure–activity relationships of this group of analogues provided evidence of the crucial role of the positive charge at the quaternary quinuclidine nitrogen atom. Moreover, the present study indicates that meta substituents, in particular halogens, on the benzyloxy substructure direct specific interactions that stabilize a desensitized conformational state of the receptor and induce silent activity

Keywords: Agonists, Cycloaddition, Nitrogen heterocycles, Receptors, Spiro compounds

Wolfenson, Haguy, Meacci, Giovanni, Liu, Shuaimin, Stachowiak, Matthew R., Iskratsch, Thomas, Ghassemi, Saba, Roca-Cusachs, Pere, Oshaughnessy, Ben, Hone, James, Sheetz, Michael P., (2016). Tropomyosin controls sarcomere-like contractions for rigidity sensing and suppressing growth on soft matrices Nature Cell Biology 18, 33-42

Cells test the rigidity of the extracellular matrix by applying forces to it through integrin adhesions. Recent measurements show that these forces are applied by local micrometre-scale contractions, but how contraction force is regulated by rigidity is unknown. Here we performed high temporal- and spatial-resolution tracking of contractile forces by plating cells on sub-micrometre elastomeric pillars. We found that actomyosin-based sarcomere-like contractile units (CUs) simultaneously moved opposing pillars in net steps of [sim]2.5[thinsp]nm, independent of rigidity. What correlated with rigidity was the number of steps taken to reach a force level that activated recruitment of [alpha]-actinin to the CUs. When we removed actomyosin restriction by depleting tropomyosin 2.1, we observed larger steps and higher forces that resulted in aberrant rigidity sensing and growth of non-transformed cells on soft matrices. Thus, we conclude that tropomyosin 2.1 acts as a suppressor of growth on soft matrices by supporting proper rigidity sensing.

Keywords: Cell adhesion, Mechanotransduction

Parra-Cabrera, C., Samitier, J., Homs-Corbera, A., (2016). Multiple biomarkers biosensor with just-in-time functionalization: Application to prostate cancer detection Biosensors and Bioelectronics 77, 1192-1200

We present a novel lab-on-a-chip (LOC) device for the simultaneous detection of multiple biomarkers using simple voltage measurements. The biosensor functionalization is performed in-situ, immediately before its use, facilitating reagents storage and massive devices fabrication. Sensitivity, limit of detection (LOD) and limit of quantification (LOQ) are tunable depending on the in-chip flown sample volumes. As a proof-of-concept, the system has been tested and adjusted to quantify two proteins found in blood that are susceptible to be used combined, as a screening tool, to diagnose prostate cancer (PCa): prostate-specific antigen (PSA) and spondin-2 (SPON2). This combination of biomarkers has been reported to be more specific for PCa diagnostics than the currently accepted but rather controversial PSA indicator. The range of detection for PSA and SPON2 could be adjusted to the clinically relevant range of 1 to 10. ng/ml. The system was tested for specificity to the evaluated biomarkers. This multiplex system can be modified and adapted to detect a larger quantity of biomarkers, or different ones, of relevance to other specific diseases.

Keywords: Adjustable sensing, Impedance measurements, In situ functionalization, Microfluidics, Prostate specific antigen, Self-assembled monolayers

Noguera-Ortega, E., Rabanal, R. M., Secanella-Fandos, S., Torrents, E., Luquin, M., Julián, E., (2016). Gamma-irradiated mycobacteria enhance survival in bladder tumor bearing mice although less efficaciously than live mycobacteria Journal of Urology , 195, (1), 198-205

Purpose γ Irradiated Mycobacterium bovis bacillus Calmette-Guérin has shown in vitro and ex vivo antitumor activity. However, to our knowledge the potential antitumor capacity has not been demonstrated in vivo. We studied the in vivo potential of γ irradiated bacillus Calmette-Guérin and γ irradiated M. brumae, a saprophytic mycobacterium that was recently described as an immunotherapeutic agent. Materials and Methods The antitumor capacity of γ irradiated M. brumae was first investigated by analyzing the in vitro inhibition of bladder tumor cell proliferation and the ex vivo cytotoxic effect of M. brumae activated peripheral blood cells. The effect of γ irradiated M. brumae or bacillus Calmette-Guérin intravesical treatment was then compared to treatment with live mycobacteria in the orthotopic murine model of bladder cancer. Results Nonviable M. brumae showed a capacity to inhibit in vitro bladder cancer cell lines similar to that of live mycobacteria. However, its capacity to induce cytokine production was decreased compared to that of live M. brumae. γ Irradiated M. brumae could activate immune cells to inhibit tumor cell growth, although to a lesser extent than live mycobacteria. Finally, intravesical treatment with γ irradiated M. brumae or bacillus Calmette-Guérin significantly increased survival with respect to that of nontreated tumor bearing mice. Both γ irradiated mycobacteria showed lower survival rates than those of live mycobacteria but the minor efficacy of γ irradiated vs live mycobacteria was only significant for bacillus Calmette-Guérin. Conclusions Our results show that although γ irradiated mycobacteria is less efficacious than live mycobacteria, it induces an antitumor effect in vivo, avoiding the possibility of further mycobacterial infections.

Keywords: BCG vaccine, Gamma rays, Immunotherapy, Mycobacterium, Urinary bladder neoplasms

Garcia-Calero, Elena, Botella-Lopez, Arancha, Bahamonde, Olga, Perez-Balaguer, Ariadna, Martinez, Salvador, (2016). FoxP2 protein levels regulate cell morphology changes and migration patterns in the vertebrate developing telencephalon Brain Structure and Function , 221, (6), 2905-2917

In the mammalian telencephalon, part of the progenitor cells transition from multipolar to bipolar morphology as they invade the mantle zone. This associates with changing patterns of radial migration. However, the molecules implicated in these morphology transitions are not well known. In the present work, we analyzed the function of FoxP2 protein in this process during telencephalic development in vertebrates. We analyzed the expression of FoxP2 protein and its relation with cell morphology and migratory patterns in mouse and chicken developing striatum. We observed FoxP2 protein expressed in a gradient from the subventricular zone to the mantle layer in mice embryos. In the FoxP2 low domain cells showed multipolar migration. In the striatal mantle layer where FoxP2 protein expression is higher, cells showed locomoting migration and bipolar morphology. In contrast, FoxP2 showed a high and homogenous expression pattern in chicken striatum, thus bipolar morphology predominated. Elevation of FoxP2 in the striatal subventricular zone by in utero electroporation promoted bipolar morphology and impaired multipolar radial migration. In mouse cerebral cortex we obtained similar results. FoxP2 promotes transition from multipolar to bipolar morphology by means of gradiental expression in mouse striatum and cortex. Together these results indicate a role of FoxP2 differential expression in cell morphology control of the vertebrate telencephalon.

Keywords: Radial migration, Bipolar morphology, Striatum, Cortex

Stanton, Morgan M., Simmchen, Juliane, Ma, Xing, Miguel-López, Albert, Sánchez, Samuel, (2016). Biohybrid Janus motors driven by Escherichia coli Advanced Materials Interfaces , 3, (2), 1500505

There has been a significant interest in the development of microswimmers for medical drug and cargo delivery, but the majority of current micromotors rely on toxic fuel sources and materials in their design making them irrelevant for biomedical applications. Bacteria represent an excellent motor alternative, as they are powered using their surrounding biological fluids. For a motile, biohybrid swimmer, Escherichia coli (E. coli) are integrated onto metal capped, polystyrene (PS) Janus particles. Fabrication of the biohybrid is rapid and simple for a microswimmer capable of magnetic guidance and ferrying an anticancer agent. Cell adhesion is regulated as E. coli adheres only to the particle's metal caps allowing the PS surface to be utilized for drug attachment, creating a multifunctional system. E. coli adhesion is investigated on multiple metal caps (Pt, Fe, Ti, or Au) and displays a strong preference to attach to Pt surfaces over other metals. Surface hydrophobicity and surface charge are examined to interpret the cell specific adhesion on the Janus particles. The dual capability of the biohybrid to have guided cell adhesion and localized drug attachment allows the swimmer to have multiple applications for biomedical microswimmers, future bacteria-interface systems, and micro-biorobots.

Keywords: Bacteria adhesion, Biohybrids, Escherichia coli, Janus particles, Microswimmers

Tahirbegi, I.B., Pardo, W.A., Alvira, M., Mir, M., Samitier, J., (2016). Amyloid Aβ 42, a promoter of magnetite nanoparticle formation in Alzheimer's disease Nanotechnology 27, (46), 465102

The accumulation of iron oxides - mainly magnetite - with amyloid peptide is a key process in the development of Alzheimer's disease (AD). However, the mechanism for biogeneration of magnetite inside the brain of someone with AD is still unclear. The iron-storing protein ferritin has been identified as the main magnetite-storing molecule. However, accumulations of magnetite in AD are not correlated with an increase in ferritin, leaving this question unresolved. Here we demonstrate the key role of amyloid peptide Aβ 42, one of the main hallmarks of AD, in the generation of magnetite nanoparticles in the absence of ferritin. The capacity of amyloid peptide to bind and concentrate iron hydroxides, the basis for the formation of magnetite, benefits the spontaneous synthesis of these nanoparticles, even under unfavorable conditions for their formation. Using scanning and transmission electron microscopy, electron energy loss spectroscopy and magnetic force microscopy we characterized the capacity of amyloid peptide Aβ 42 to promote magnetite formation.

Keywords: Alzheimer disease (AD), amyloid peptide Ab42, magnetite nanoparticle, metallobiomolecule, iron oxide, neurodegenerative brain diseases

Lozano-Garcia, M., Fiz, J. A., Jané, R., (2016). Performance evaluation of the Hilbert–Huang transform for respiratory sound analysis and its application to continuous adventitious sound characterization Signal Processing , 120, 99-116

Abstract The use of the Hilbert–Huang transform in the analysis of biomedical signals has increased during the past few years, but its use for respiratory sound (RS) analysis is still limited. The technique includes two steps: empirical mode decomposition (EMD) and instantaneous frequency (IF) estimation. Although the mode mixing (MM) problem of EMD has been widely discussed, this technique continues to be used in many RS analysis algorithms. In this study, we analyzed the MM effect in RS signals recorded from 30 asthmatic patients, and studied the performance of ensemble EMD (EEMD) and noise-assisted multivariate EMD (NA-MEMD) as means for preventing this effect. We propose quantitative parameters for measuring the size, reduction of MM, and residual noise level of each method. These parameters showed that EEMD is a good solution for MM, thus outperforming NA-MEMD. After testing different IF estimators, we propose Kay׳s method to calculate an EEMD-Kay-based Hilbert spectrum that offers high energy concentrations and high time and high frequency resolutions. We also propose an algorithm for the automatic characterization of continuous adventitious sounds (CAS). The tests performed showed that the proposed EEMD-Kay-based Hilbert spectrum makes it possible to determine CAS more precisely than other conventional time-frequency techniques.

Keywords: Hilbert–Huang transform, Ensemble empirical mode decomposition, Instantaneous frequency, Respiratory sounds, Continuous adventitious sounds

Páez-Avilés, C., Juanola-Feliu, E., Punter-Villagrasa, J., Del Moral Zamora, B., Homs-Corbera, A., Colomer-Farrarons, J., Miribel-Català , P. L., Samitier, J., (2016). Combined dielectrophoresis and impedance systems for bacteria analysis in microfluidic on-chip platforms Sensors 16, (9), 1514

Bacteria concentration and detection is time-consuming in regular microbiology procedures aimed to facilitate the detection and analysis of these cells at very low concentrations. Traditional methods are effective but often require several days to complete. This scenario results in low bioanalytical and diagnostic methodologies with associated increased costs and complexity. In recent years, the exploitation of the intrinsic electrical properties of cells has emerged as an appealing alternative approach for concentrating and detecting bacteria. The combination of dielectrophoresis (DEP) and impedance analysis (IA) in microfluidic on-chip platforms could be key to develop rapid, accurate, portable, simple-to-use and cost-effective microfluidic devices with a promising impact in medicine, public health, agricultural, food control and environmental areas. The present document reviews recent DEP and IA combined approaches and the latest relevant improvements focusing on bacteria concentration and detection, including selectivity, sensitivity, detection time, and conductivity variation enhancements. Furthermore, this review analyses future trends and challenges which need to be addressed in order to successfully commercialize these platforms resulting in an adequate social return of public-funded investments.

Keywords: Bacteria, Dielectrophoresis, Impedance, Microfluidics, On-chip

Coelho, N. M., Llopis-Hernández, V., Salmerón-Sánchez, M., Altankov, G., (2016). Dynamic reorganization and enzymatic remodeling of type IV collagen at cell–biomaterial interface Advances in Protein Chemistry and Structural Biology (ed. Christo, Z. Christov), Academic Press (San Diego, USA) 105, 81-104

Abstract Vascular basement membrane remodeling involves assembly and degradation of its main constituents, type IV collagen (Col IV) and laminin, which is critical during development, angiogenesis, and tissue repair. Remodeling can also occur at cell–biomaterials interface altering significantly the biocompatibility of implants. Here we describe the fate of adsorbed Col IV in contact with endothelial cells adhering on positively charged NH2 or hydrophobic CH3 substrata, both based on self-assembly monolayers (SAMs) and studied alone or mixed in different proportions. AFM studies revealed distinct pattern of adsorbed Col IV, varying from single molecular deposition on pure NH2 to network-like assembly on mixed SAMs, turning to big globular aggregates on bare CH3. Human umbilical endothelial cells (HUVECs) interact better with Col IV adsorbed as single molecules on NH2 surface and readily rearrange it in fibril-like pattern that coincide with secreted fibronectin fibrils. The cells show flattened morphology and well-developed focal adhesion complexes that are rich on phosphorylated FAK while expressing markedly low pericellular proteolytic activity. Conversely, on hydrophobic CH3 substrata HUVECs showed abrogated spreading and FAK phosphorylation, combined with less reorganization of the aggregated Col IV and significantly increased proteolytic activity. The later involves both MMP-2 and MMP-9, as measured by zymography and FITC-Col IV release. The mixed SAMs support intermediate remodeling activity. Taken together these results show that chemical functionalization combined with Col IV preadsorption provides a tool for guiding the endothelial cells behavior and pericellular proteolytic activity, events that strongly affect the fate of cardiovascular implants.

Keywords: Type IV collagen, Adsorption, Remodeling, Pericellular proteolysis, Reorganization, Substratum chemistry, CH3 and NH2 groups, Self-assembly monolayers

Lagunas, Anna, Martinez, Elena, Samitier, Josep, (2015). Surface-bound molecular gradients for the high throughput screening of cell responses Frontiers in Bioengineering and Biotechnology 3, Article 132

Chemical gradient surfaces are described as surfaces with a gradually varying composition along their length. Continuous chemical gradients have recently been proposed as alternative to discrete microarrays for the high throughput screening of the effects of ligand concentration in cells. Here we review some of the most recent examples in which gradients have been used to evaluate the effect of a varying ligand concentration in cell adhesion, morphology, growth and differentiation of cells, including some of our recent findings. They show the importance of the organization of ligands at the nanoscale, which is highlighted by abrupt changes in cell behavior at critical concentration thresholds.

Keywords: Cell Adhesion, Cell Differentiation, Cell growth, Cell morphology, Molecular gradient

Crosas-Molist, E., Meirelles, T., López-Luque, J., Serra-Peinado, C., Selva, J., Caja, L., Gorbenko Del Blanco, D., Uriarte, J. J., Bertran, E., Mendizábal, Y., Hernández, V., García-Calero, C., Busnadiego, O., Condom, E., Toral, D., Castellà, M., Forteza, A., Navajas, D., Sarri, E., Rodríguez-Pascual, F., Dietz, H. C., Fabregat, I., Egea, G., (2015). Vascular smooth muscle cell phenotypic changes in patients with marfan syndrome Arteriosclerosis, Thrombosis, and Vascular Biology , 35, (4), 960-972

Objective - Marfan's syndrome is characterized by the formation of ascending aortic aneurysms resulting from altered assembly of extracellular matrix microfibrils and chronic tissue growth factor (TGF)-β signaling. TGF-β is a potent regulator of the vascular smooth muscle cell (VSMC) phenotype. We hypothesized that as a result of the chronic TGF-β signaling, VSMC would alter their basal differentiation phenotype, which could facilitate the formation of aneurysms. This study explores whether Marfan's syndrome entails phenotypic alterations of VSMC and possible mechanisms at the subcellular level. Approach and Results - Immunohistochemical and Western blotting analyses of dilated aortas from Marfan patients showed overexpression of contractile protein markers (α-smooth muscle actin, smoothelin, smooth muscle protein 22 alpha, and calponin-1) and collagen I in comparison with healthy aortas. VSMC explanted from Marfan aortic aneurysms showed increased in vitro expression of these phenotypic markers and also of myocardin, a transcription factor essential for VSMC-specific differentiation. These alterations were generally reduced after pharmacological inhibition of the TGF-β pathway. Marfan VSMC in culture showed more robust actin stress fibers and enhanced RhoA-GTP levels, which was accompanied by increased focal adhesion components and higher nuclear localization of myosin-related transcription factor A. Marfan VSMC and extracellular matrix measured by atomic force microscopy were both stiffer than their respective controls. Conclusions - In Marfan VSMC, both in tissue and in culture, there are variable TGF-β-dependent phenotypic changes affecting contractile proteins and collagen I, leading to greater cellular and extracellular matrix stiffness. Altogether, these alterations may contribute to the known aortic rigidity that precedes or accompanies Marfan's syndrome aneurysm formation.

Keywords: Actin, Aortic aneurysms, Aortic stiffness, Extracellular matrix, Focal adhesion, Myocardin, RhoA, TGF-β

Abadías, Clara, Serés, Carme, Torrent-Burgués, J., (2015). AFM in peak force mode applied to worn siloxane-hydrogel contact lenses Colloids and Surfaces B: Biointerfaces 128, 61-66

The objective of this work is to apply Atomic Force Microscopy in Peak Force mode to obtain topographic characteristics (mean roughness, root-mean-square roughness, skewness and kurtosis) and mechanical characteristics (adhesion, elastic modulus) of Siloxane-Hydrogel Soft Contact Lenses (CLs) of two different materials, Lotrafilcon B of Air Optix (AO) and Asmofilcon A of PremiO (P), after use (worn CLs). Thus, the results obtained with both materials will be compared, as well as the changes produced by the wear at a nanoscopic level. The results show significant changes in the topographic and mechanical characteristics of the CLs, at a nanoscopic level, due to wear. The AO CL show values of the topographic parameters lower than those of the P CL after wear, which correlates with a better comfort qualification given to the former by the wearers. A significant correlation has also been obtained between the adhesion values found after the use of the CLs with tear quality tests, both break-up-time and Schirmer.

Keywords: Adhesion, Atomic force microscopy-peak force mode, Surface topography, Worn siloxane-hydrogel contact lenses, Young modulus

Mrkonji, Garcia-Elias, A., Pardo-Pastor, C., Bazellières, E., Trepat, X., Vriens, J., Ghosh, D., Voets, T., Vicente, R., Valverde, M. A., (2015). TRPV4 participates in the establishment of trailing adhesions and directional persistence of migrating cells Pflugers Archiv European Journal of Physiology , 467, (10), 2107-2119

Calcium signaling participates in different cellular processes leading to cell migration. TRPV4, a non-selective cation channel that responds to mechano-osmotic stimulation and heat, is also involved in cell migration. However, the mechanistic involvement of TRPV4 in cell migration is currently unknown. We now report that expression of the mutant channel TRPV4-121AAWAA (lacking the phosphoinositide-binding site 121KRWRK125 and the response to physiological stimuli) altered HEK293 cell migration. Altered migration patterns included periods of fast and persistent motion followed by periods of stalling and turning, and the extension of multiple long cellular protrusions. TRPV4-WT overexpressing cells showed almost complete loss of directionality with frequent turns, no progression, and absence of long protrusions. Traction microscopy revealed higher tractions forces in the tail of TRPV4-121AAWAA than in TRPV4-WT expressing cells. These results are consistent with a defective and augmented tail retraction in TRPV4-121AAWAA- and TRPV4-WT-expressing cells, respectively. The activity of calpain, a protease implicated in focal adhesion (FA) disassembly, was decreased in TRPV4-121AAWAA compared with TRPV4-WT-expressing cells. Consistently, larger focal adhesions were seen in TRPV4-121AAWAA compared with TRPV4-WT-expressing HEK293 cells, a result that was also reproduced in T47D and U87 cells. Similarly, overexpression of the pore-dead mutant TRPV4-M680D resumed the TRPV4-121AAWAA phenotype presenting larger FA. The migratory phenotype obtained in HEK293 cells overexpressing TRPV4-121AAWAA was mimicked by knocking-down TRPC1, a cationic channel that participates in cell migration. Together, our results point to the participation of TRPV4 in the dynamics of trailing adhesions, a function that may require the interplay of TRPV4 with other cation channels or proteins present at the FA sites.

Keywords: Calcium, Calpain, Focal adhesion, Migration, Traction forces, TRPV4

Estévez, M., Martínez, Elena, Yarwood, S. J., Dalby, M. J., Samitier, J., (2015). Adhesion and migration of cells responding to microtopography Journal of Biomedical Materials Research - Part A , 103, (5), 1659-1668

It is known that cells respond strongly to microtopography. However, cellular mechanisms of response are unclear. Here, we study wild-type fibroblasts responding to 25 μm2 posts and compare their response to that of FAK-/- fibroblasts and fibroblasts with PMA treatment to stimulate protein kinase C (PKC) and the small g-protein Rac. FAK knockout cells modulated adhesion number and size in a similar way to cells on topography; that is, they used more, smaller adhesions, but migration was almost completely stalled demonstrating the importance of FAK signaling in contact guidance and adhesion turnover. Little similarity, however, was observed to PKC stimulated cells and cells on the topography. Interestingly, with PKC stimulation the cell nuclei became highly deformable bringing focus on these surfaces to the study of metastasis. Surfaces that aid the study of cellular migration are important in developing understanding of mechanisms of wound healing and repair in aligned tissues such as ligament and tendon.

Keywords: Adhesion, Cell migration, Cell morphology, Focal adhesion kinase, Microstructures

Perea-Gil, I., Uriarte, J. J., Prat-Vidal, C., Gálvez-Montón, C., Roura, S., Llucià-Valldeperas, A., Soler-Botija, C., Farré, R., Navajas, D., Bayes-Genis, A., (2015). In vitro comparative study of two decellularization protocols in search of an optimal myocardial scaffold for recellularization American Journal of Translational Research , 7, (3), 558-573

Introduction. Selection of a biomaterial-based scaffold that mimics native myocardial extracellular matrix (ECM) architecture can facilitate functional cell attachment and differentiation. Although decellularized myocardial ECM accomplishes these premises, decellularization processes may variably distort or degrade ECM structure. Materials and methods. Two decellularization protocols (DP) were tested on porcine heart samples (epicardium, mid myocardium and endocardium). One protocol, DP1, was detergent-based (SDS and Triton X-100), followed by DNase I treatment. The other protocol, DP2, was focused in trypsin and acid with Triton X-100 treatments. Decellularized myocardial scaffolds were reseeded by embedding them in RAD16-I peptidic hydrogel with adipose tissue-derived progenitor cells (ATDPCs). Results. Both protocols yielded acellular myocardial scaffolds (~82% and ~94% DNA reduction for DP1 and DP2, respectively). Ultramicroscopic assessment of scaffolds was similar for both protocols and showed filamentous ECM with preserved fiber disposition and structure. DP1 resulted in more biodegradable scaffolds (P = 0.04). Atomic force microscopy revealed no substantial ECM stiffness changes post-decellularization compared to native tissue. The Young’s modulus did not differ between heart layers (P = 0.69) or decellularization protocols (P = 0.15). After one week, recellularized DP1 scaffolds contained higher cell density (236 ± 106 and 98 ± 56 cells/mm2 for recellularized DP1 and DP2 scaffolds, respectively; P = 0.04). ATDPCs in both DP1 and DP2 scaffolds expressed the endothelial marker isolectin B4, but only in the DP1 scaffold ATDPCs expressed the cardiac markers GATA4, connexin43 and cardiac troponin T. Conclusions. In our hands, DP1 produced myocardial scaffolds with higher cell repopulation and promotes ATDPCs expression of endothelial and cardiomyogenic markers.

Keywords: Acellular myocardial scaffold, Adipose tissue-derived progenitor cells, Decellularization protocols, Extracellular matrix, Myocardial infarction, Recellularization

Gumí-Audenis, B., Carlà, F., Vitorino, M. V., Panzarella, A., Porcar, L., Boilot, M., Guerber, S., Bernard, P., Rodrigues, M. S., Sanz, F., Giannotti, M. I., Costa, L., (2015). Custom AFM for X-ray beamlines: in situ biological investigations under physiological conditions Journal of Synchrotron Radiation , 22, 1364-1371

A fast atomic force microscope (AFM) has been developed that can be installed as a sample holder for grazing-incidence X-ray experiments at solid/gas or solid/liquid interfaces. It allows a wide range of possible investigations, including soft and biological samples under physiological conditions (hydrated specimens). The structural information obtained using the X-rays is combined with the data gathered with the AFM (morphology and mechanical properties), providing a unique characterization of the specimen and its dynamics in situ during an experiment. In this work, lipid monolayers and bilayers in air or liquid environment have been investigated by means of AFM, both with imaging and force spectroscopy, and X-ray reflectivity. In addition, this combination allows the radiation damage induced by the beam on the sample to be studied, as has been observed on DOPC and DPPC supported lipid bilayers under physiological conditions.

Keywords: In situ atomic force microscopy, Grazing-incidence scattering and reflectivity, Radiation damage, Model lipid membranes

Rajasekaran, Vijaykumar, Aranda, Joan, Casals, Alicia, Pons, Jose L., (2015). An adaptive control strategy for postural stability using a wearable robot Robotics and Autonomous Systems , 73, 16-23

Abstract Wearable robots are expected to expand the use of robotics in rehabilitation since they can widen the assistance application context. An important aspect of a rehabilitation therapy, in terms of lower extremity assistance, is balance control. In this article, we propose and evaluate an adaptive control strategy for robotic rehabilitation therapies to guarantee static stability using a wearable robot. Postural balance control can be implemented either acting on the hip, on the ankle joint or on both, depending on the kind of perturbation acting on the subject: internal or external. Internal perturbations can be produced by any voluntary movement of the body, such as bending the trunk. External perturbations, in the form of an impact force, are applied by the exoskeleton without any prior notice to observe the proactive response of the subject. We have used a 6 degree of freedom planar lower limb exoskeleton, H1, to perform this analysis. The developed control strategy has been designed to provide the necessary assistance, related to balance recovery and postural stability, under the “Assist-as-needed” paradigm. The interaction forces between orthosis and subject are monitored, as they play a relevant role in the definition of assistive and resistive movements to be applied to the joints. The proposed method has been tested with 5 healthy subjects in presence of internal and external disturbances. The results demonstrate that knowing the stability limit of each subject, in combination with a therapeutically selected scaling factor, the proposed adaptive control helps in providing an effective assistance in therapy. This method is efficient in handling the individual and combined effect of external perturbations acting on any joint movements.

Keywords: Exoskeleton controls, Postural stability, Balance controls, Adaptive control

Keremidarska, M., Gugutkov, D., Altankov, G., Krasteva, N., (2015). Impact of electrospun nanofibres orientation on mesenchymal stem cell adhesion and morphology Comptes Rendus de L'Academie Bulgare des Sciences , 68, (10), 1271-1276

Electrospun nanofibrous materials mimicking the architecture of native extracellular matrix (ECM) hold great promise as scaffolds in tissue engineering. In order to optimize the properties of nanofibrous scaffolds it is important to understand the impact of fibres’ organization on cell behaviour. Herein, we investigated the effect of nanofibres (NFs) alignment on human adipose-derived mesenchymal stem cells (hAD-MSCs) adhesion and morphology. Electrospun composite fibrinogen/poly-lactic acid (FNG/PLA) NF scaffolds with same composition and comparable fibre size were fabricated into randomly oriented and aligned configuration and stem cells adhesion was characterized by the meaning of overall cell morphology, actin cytoskeleton organization and expression of molecules, involved in the development of focal adhesion complexes. We found that hAD-MSCs altered their morphology, actin cytoskeleton and cell attachment in accordance with nanofibre orientation while cell spreading, focal adhesions and expression of β1 and αNintegrin receptors were not influenced significantly by fibre orientation. These results confirmed that fibre alignment of scaffold guide cellular arrangement and could be beneficial for stem differentiation and therefore for the successful scaffolds development if its contact guidance coincided with the cell shape and cytoskeletal tension.

Keywords: Electrospinning, Fibrinogen/polylactic acid hybrid nanofibres, Human adipose-derived stem cells

Rajasekaran, V., Aranda, J., Casals, A., (2015). Compliant gait assistance triggered by user intention Engineering in Medicine and Biology Society (EMBC) 37th Annual International Conference of the IEEE , IEEE (Milan, Italy) , 3885-3888

An automatic gait initialization strategy based on user intention sensing in the context of rehabilitation with a lower-limb wearable robot is proposed and evaluated. The proposed strategy involves monitoring the human-orthosis interaction torques and initial position deviation to determine the gait initiation instant and to modify orthosis operation for gait assistance, when needed. During gait, the compliant control algorithm relies on the adaptation of the joints' stiffness in function of their interaction torques and their deviation from the desired trajectories, while maintaining the dynamic stability. As a reference input, the average of a set of recorded gaits obtained from healthy subjects is used. The algorithm has been tested with five healthy subjects showing its efficient behavior in initiating the gait and maintaining the equilibrium while walking in presence of external forces. The work is performed as a preliminary study to assist patients suffering from incomplete Spinal cord injury and Stroke.

Keywords: Biomedical monitoring, Exoskeletons, Joints, Knee, Legged locomotion, Trajectory, Exoskeleton, adaptive control, gait assistance, gait initiation, rehabilitation, wearable robot

Comelles, J., Hortigüela, V., Martínez, Elena, Riveline, D., (2015). Methods for rectifying cell motions in vitro: Breaking symmetry using microfabrication and microfluidics Methods in Cell Biology - Biophysical Methods in Cell Biology (ed. Wilson, L., Tran, P.), Academic Press (Santa Barbara, USA) 125, 437-452

Cell motility is an important phenomenon in cell biology, developmental biology, and cancer. Here we report methods that we designed to identify and characterize external factors which direct cell motions by breaking locally the symmetry. We used microfabrication and microfluidics techniques to impose and combine mechanical and chemical cues to moving fibroblasts. Gradients can thereby be engineered at the cellular scale and this approach has allowed to disentangle roles of the nucleus and protrusion activity in setting cell directions.

Keywords: Adhesion, Biological physics, Cell motility, Gradient, Ratchet

Rajasekaran, V., Aranda, J., Casals, A., (2015). User intention driven adaptive gait assistance using a wearable exoskeleton Robot 2015: Second Iberian Robotics Conference (ed. Paulo Reis, L., Paulo Moreira, A., Lima, P. U., Montano, L., Muñoz-Martinez, V.), Springer International (Lausanne, Switzerland) 418, 289-301

A user intention based rehabilitation strategy for a lower-limb wearable robot is proposed and evaluated. The control strategy, which involves monitoring the human-orthosis interaction torques, determines the gait initiation instant and modifies orthosis operation for gait assistance, when needed. Orthosis operation is classified as assistive or resistive in function of its evolution with respect to a normal gait pattern. The control algorithm relies on the adaptation of the joints’ stiffness in function of their interaction torques and their deviation from the desired trajectories. An average of recorded gaits obtained from healthy subjects is used as reference input. The objective of this work is to develop a control strategy that can trigger the gait initiation from the user’s intention and maintain the dynamic stability, using an efficient real-time stiffness adaptation for multiple joints, simultaneously maintaining their synchronization. The algorithm has been tested with five healthy subjects showing its efficient behavior in initiating the gait and maintaining the equilibrium while walking in presence of external forces. The work is performed as a preliminary study to assist patients suffering from incomplete Spinal cord injury and Stroke.

Keywords: Adaptive control, Exoskeleton, Gait assistance, Gait initiation, Wearable robot

Cuervo, A., Dans, P. D., Carrascosa, J. L., Orozco, M., Gomila, G., Fumagalli, L., (2014). Direct measurement of the dielectric polarization properties of DNA Proceedings of the National Academy of Sciences of the United States of America 111, (35), E3624-E3630

The electric polarizability of DNA, represented by the dielectric constant, is a key intrinsic property that modulates DNA interaction with effector proteins. Surprisingly, it has so far remained unknown owing to the lack of experimental tools able to access it. Here, we experimentally resolved it by detecting the ultraweak polarization forces of DNA inside single T7 bacteriophages particles using electrostatic force microscopy. In contrast to the common assumption of low-polarizable behavior like proteins (εr ~ 2–4), we found that the DNA dielectric constant is ~ 8, considerably higher than the value of ~ 3 found for capsid proteins. State-of-the-art molecular dynamic simulations confirm the experimental findings, which result in sensibly decreased DNA interaction free energy than normally predicted by Poisson–Boltzmann methods. Our findings reveal a property at the basis of DNA structure and functions that is needed for realistic theoretical descriptions, and illustrate the synergetic power of scanning probe microscopy and theoretical computation techniques.

Keywords: Atomic force microscopy, Atomistic simulations, DNA packaging, DNA-ligand binding, Poisson-Boltzmann equation, capsid protein, DNA, double stranded DNA, amino acid composition, article, atomic force microscopy, bacteriophage, bacteriophage T7, dielectric constant, dipole, DNA binding, DNA packaging, DNA structure, electron microscopy, ligand binding, nonhuman, polarization, priority journal, protein analysis, protein DNA interaction, scanning probe microscopy, static electricity, virion, virus capsid, virus particle, atomic force microscopy, atomistic simulations, DNA packaging, DNA-ligand binding, Poisson-Boltzmann equation, Bacteriophage T7, Capsid, Cations, Dielectric Spectroscopy, DNA, DNA, Viral, DNA-Binding Proteins, Electrochemical Techniques, Ligands, Microscopy, Atomic Force, Models, Chemical, Nuclear Proteins

Lagunas, A., Garcia, A., Artés, J. M., Vida, Y., Collado, D., Pérez-Inestrosa, E., Gorostiza, P., Claros, S., Andrades, J. A., Samitier, J., (2014). Large-scale dendrimer-based uneven nanopatterns for the study of local arginine-glycine-aspartic acid (RGD) density effects on cell adhesion Nano Research , 7, (3), 399-409

Cell adhesion processes are governed by the nanoscale arrangement of the extracellular matrix (ECM), being more affected by local rather than global concentrations of cell adhesive ligands. In many cell-based studies, grafting of dendrimers on surfaces has shown the benefits of the local increase in concentration provided by the dendritic configuration, although the lack of any reported surface characterization has limited any direct correlation between dendrimer disposition and cell response. In order to establish a proper correlation, some control over dendrimer surface deposition is desirable. Here, dendrimer nanopatterning has been employed to address arginine-glycine-aspartic acid (RGD) density effects on cell adhesion. Nanopatterned surfaces were fully characterized by atomic force microscopy (AFM), scanning tunneling microscopy (STM) and X-ray photoelectron spectroscopy (XPS), showing that tunable distributions of cell adhesive ligands on the surface are obtained as a function of the initial dendrimer bulk concentration. Cell experiments showed a clear correlation with dendrimer surface layout: Substrates presenting regions of high local ligand density resulted in a higher percentage of adhered cells and a higher degree of maturation of focal adhesions (FAs). Therefore, dendrimer nanopatterning is presented as a suitable and controlled approach to address the effect of local ligand density on cell response. Moreover, due to the easy modification of dendrimer peripheral groups, dendrimer nanopatterning can be further extended to other ECM ligands having density effects on cells.

Keywords: Arginine-glycine-aspartic acid, Atomic force microscopy, Cell adhesion, Dendrimer, Focal adhesions, Scanning tunneling microscopy

Castaño, O., Sachot, N., Xuriguera, E., Engel, E., Planell, J. A., Park, J. H., Jin, G. Z., Kim, T. H., Kim, J. H., Kim, H. W., (2014). Angiogenesis in bone regeneration: Tailored calcium release in hybrid fibrous scaffolds ACS Applied Materials & Interfaces 6, (10), 7512-7522

In bone regeneration, silicon-based calcium phosphate glasses (Bioglasses) have been widely used since the 1970s. However, they dissolve very slowly because of their high amount of Si (SiO2 > 45%). Recently, our group has found that calcium ions released by the degradation of glasses in which the job of silicon is done by just 5% of TiO2 are effective angiogenic promoters, because of their stimulation of a cell-membrane calcium sensing receptor (CaSR). Based on this, other focused tests on angiogenesis have found that Bioglasses also have the potential to be angiogenic promoters even with high contents of silicon (80%); however, their slow degradation is still a problem, as the levels of silicon cannot be decreased any lower than 45%. In this work, we propose a new generation of hybrid organically modified glasses, ormoglasses, that enable the levels of silicon to be reduced, therefore speeding up the degradation process. Using electrospinning as a faithful way to mimic the extracellular matrix (ECM), we successfully produced hybrid fibrous mats with three different contents of Si (40, 52, and 70%), and thus three different calcium ion release rates, using an ormoglass–polycaprolactone blend approach. These mats offered a good platform to evaluate different calcium release rates as osteogenic promoters in an in vivo subcutaneous environment. Complementary data were collected to complement Ca2+ release analysis, such as stiffness evaluation by AFM, ζ-potential, morphology evaluation by FESEM, proliferation and differentiation analysis, as well as in vivo subcutaneous implantations. Material and biological characterization suggested that compositions of organic/inorganic hybrid materials with a Si content equivalent to 40%, which were also those that released more calcium, were osteogenic. They also showed a greater ability to form blood vessels. These results suggest that Si-based ormoglasses can be considered an efficient tool for calcium release modulation, which could play a key role in the angiogenic promoting process.

Keywords: Biological materials, Blood vessels, Calcium, Electrospinning, Glass, Hybrid materials, Silicon oxides, Sol-gel process, Sol-gels, Angiogenesis, Biological characterization, Calcium phosphate glass, Calcium-sensing receptors, Degradation process, Extracellular matrices, Organic/inorganic hybrid materials, ormoglasses, Silicon

Pérez-Madrigal, M. M., Giannotti, M. I., Del Valle, L. J., Franco, L., Armelin, E., Puiggalí, J., Sanz, F., Alemán, C., (2014). Thermoplastic polyurethane:polythiophene nanomembranes for biomedical and biotechnological applications ACS Applied Materials & Interfaces 6, (12), 9719-9732

Nanomembranes have been prepared by spin-coating mixtures of a polythiophene (P3TMA) derivative and thermoplastic polyurethane (TPU) using 20:80, 40:60, and 60:40 TPU:P3TMA weight ratios. After structural, topographical, electrochemical, and thermal characterization, properties typically related with biomedical applications have been investigated: swelling, resistance to both hydrolytic and enzymatic degradation, biocompatibility, and adsorption of type I collagen, which is an extra cellular matrix protein that binds fibronectin favoring cell adhesion processes. The swelling ability and the hydrolytic and enzymatic degradability of TPU:P3TMA membranes increases with the concentration of P3TMA. Moreover, the degradation of the blends is considerably promoted by the presence of enzymes in the hydrolytic medium, TPU:P3TMA blends behaving as biodegradable materials. On the other hand, TPU:P3TMA nanomembranes behave as bioactive platforms stimulating cell adhesion and, especially, cell viability. Type I collagen adsorption largely depends on the substrate employed to support the nanomembrane, whereas it is practically independent of the chemical nature of the polymeric material used to fabricate the nanomembrane. However, detailed microscopy study of the morphology and topography of adsorbed collagen evidence the formation of different organizations, which range from fibrils to pseudoregular honeycomb networks depending on the composition of the nanomembrane that is in contact with the protein. Scaffolds made of electroactive TPU:P3TMA nanomembranes are potential candidates for tissue engineering biomedical applications.

Keywords: Bioactive platform, Biodegradable blend, Collaged adsorption, Scaffolds, Tissue engineering, Ultrathin films

Marques, J., Moles, E., Urbán, P., Prohens, R., Busquets, M. A., Sevrin, C., Grandfils, C., Fernàndez-Busquets, X., (2014). Application of heparin as a dual agent with antimalarial and liposome targeting activities toward Plasmodium-infected red blood cells Nanomedicine: Nanotechnology, Biology, and Medicine 10, (8), 1719-1728

Heparin had been demonstrated to have antimalarial activity and specific binding affinity for Plasmodium-infected red blood cells (pRBCs) vs. non-infected erythrocytes. Here we have explored if both properties could be joined into a drug delivery strategy where heparin would have a dual role as antimalarial and as a targeting element of drug-loaded nanoparticles. Confocal fluorescence and transmission electron microscopy data show that after 30. min of being added to living pRBCs fluorescein-labeled heparin colocalizes with the intracellular parasites. Heparin electrostatically adsorbed onto positively charged liposomes containing the cationic lipid 1,2-dioleoyl-3-trimethylammonium-propane and loaded with the antimalarial drug primaquine was capable of increasing three-fold the activity of encapsulated drug in Plasmodium falciparum cultures. At concentrations below those inducing anticoagulation of mouse blood in vivo, parasiticidal activity was found to be the additive result of the separate activities of free heparin as antimalarial and of liposome-bound heparin as targeting element for encapsulated primaquine. From the Clinical Editor: Malaria remains an enormous global public health concern. In this study, a novel functionalized heparin formulation used as drug delivery agent for primaquine was demonstrated to result in threefold increased drug activity in cell cultures, and in a murine model it was able to provide these benefits in concentrations below what would be required for anticoagulation. Further studies are needed determine if this approach is applicable in the human disease as well.

Keywords: Heparin, Liposomes, Malaria, Plasmodium, Targeted drug delivery, Heparin, Malaria, Plasmodium, Red blood cell, Targeted drug delivery, Liposomes, 1,2 dioleoyl 3 trimethylammoniopropane, fluorescein, heparin, liposome, nanoparticle, primaquine, adsorption, animal experiment, anticoagulation, antimalarial activity, Article, binding affinity, confocal microscopy, controlled study, drug targeting, encapsulation, erythrocyte, female, fluorescence microscopy, human, human cell, in vivo study, liposomal delivery, mouse, nonhuman, Plasmodium falciparum, transmission electron microscopy

Gil, V., Nocentini, S., del Río, J. A., (2014). Historical first descriptions of Cajal-Retzius cells: From pioneer studies to current knowledge Frontiers in Neuroanatomy , 8, Article 32 (9)

Santiago Ramón y Cajal developed a great body of scientific research during the last decade of 19th century, mainly between 1888 and 1892, when he published more than 30 manuscripts. The neuronal theory, the structure of dendrites and spines, and fine microscopic descriptions of numerous neural circuits are among these studies. In addition, numerous cell types (neuronal and glial) were described by Ramón y Cajal during this time using this "reazione nera" or Golgi method. Among these neurons were the special cells of the molecular layer of the neocortex. These cells were also termed Cajal cells or Retzius cells by other colleagues. Today these cells are known as Cajal-Retzius cells. From the earliest description, several biological aspects of these fascinating cells have been analyzed (e.g., cell morphology, physiological properties, origin and cellular fate, putative function during cortical development, etc). In this review we will summarize in a temporal basis the emerging knowledge concerning this cell population with specific attention the pioneer studies of Santiago Ramón y Cajal.

Keywords: Calretinin, Cortical hem, Neocortical development, Pioneer neurons, Radial glia, Reelin

Melo, E., Cárdenes, N., Garreta, E., Luque, T., Rojas, M., Navajas, D., Farré, R., (2014). Inhomogeneity of local stiffness in the extracellular matrix scaffold of fibrotic mouse lungs Journal of the Mechanical Behavior of Biomedical Materials , 37, 186-195

Lung disease models are useful to study how cell engraftment, proliferation and differentiation are modulated in lung bioengineering. The aim of this work was to characterize the local stiffness of decellularized lungs in aged and fibrotic mice. Mice (2- and 24-month old; 14 of each) with lung fibrosis (N=20) and healthy controls (N=8) were euthanized after 11 days of intratracheal bleomycin (fibrosis) or saline (controls) infusion. The lungs were excised, decellularized by a conventional detergent-based (sodium-dodecyl sulfate) procedure and slices of the acellular lungs were prepared to measure the local stiffness by means of atomic force microscopy. The local stiffness of the different sites in acellular fibrotic lungs was very inhomogeneous within the lung and increased according to the degree of the structural fibrotic lesion. Local stiffness of the acellular lungs did not show statistically significant differences caused by age. The group of mice most affected by fibrosis exhibited local stiffness that were ~2-fold higher than in the control mice: from 27.2±1.64 to 64.8±7.1. kPa in the alveolar septa, from 56.6±4.6 to 99.9±11.7. kPa in the visceral pleura, from 41.1±8.0 to 105.2±13.6. kPa in the tunica adventitia, and from 79.3±7.2 to 146.6±28.8. kPa in the tunica intima. Since acellular lungs from mice with bleomycin-induced fibrosis present considerable micromechanical inhomogeneity, this model can be a useful tool to better investigate how different degrees of extracellular matrix lesion modulate cell fate in the process of organ bioengineering from decellularized lungs.

Keywords: Ageing, Atomic force microscopy, Decellularization, Lung fibrosis, Tissue engineering, Atomic force microscopy, Biological organs, Peptides, Sodium dodecyl sulfate, Sodium sulfate, Tissue engineering, Ageing, Decellularization, Extracellular matrices, Healthy controls, Inhomogeneities, Lung fibrosis, Micro-mechanical, Statistically significant difference, Mammals, bleomycin, adventitia, animal experiment, animal model, article, atomic force microscopy, bleomycin-induced pulmonary fibrosis, cell fate, controlled study, extracellular matrix, female, intima, lung alveolus, lung fibrosis, lung mechanics, mechanical probe, microenvironment, mouse, nonhuman, pleura, priority journal, rigidity, tissue engineering

Uriarte, J. J., Nonaka, P. N., Campillo, N., Palma, R. K., Melo, E., de Oliveira, L. V. F., Navajas, D., Farré, R., (2014). Mechanical properties of acellular mouse lungs after sterilization by gamma irradiation Journal of the Mechanical Behavior of Biomedical Materials , 40, 168-177

Lung bioengineering using decellularized organ scaffolds is a potential alternative for lung transplantation. Clinical application will require donor scaffold sterilization. As gamma-irradiation is a conventional method for sterilizing tissue preparations for clinical application, the aim of this study was to evaluate the effects of lung scaffold sterilization by gamma irradiation on the mechanical properties of the acellular lung when subjected to the artificial ventilation maneuvers typical within bioreactors. Twenty-six mouse lungs were decellularized by a sodium dodecyl sulfate detergent protocol. Eight lungs were used as controls and 18 of them were submitted to a 31kGy gamma irradiation sterilization process (9 kept frozen in dry ice and 9 at room temperature). Mechanical properties of acellular lungs were measured before and after irradiation. Lung resistance (RL) and elastance (EL) were computed by linear regression fitting of recorded signals during mechanical ventilation (tracheal pressure, flow and volume). Static (Est) and dynamic (Edyn) elastances were obtained by the end-inspiratory occlusion method. After irradiation lungs presented higher values of resistance and elastance than before irradiation: RL increased by 41.1% (room temperature irradiation) and 32.8% (frozen irradiation) and EL increased by 41.8% (room temperature irradiation) and 31.8% (frozen irradiation). Similar increases were induced by irradiation in Est and Edyn. Scanning electron microscopy showed slight structural changes after irradiation, particularly those kept frozen. Sterilization by gamma irradiation at a conventional dose to ensure sterilization modifies acellular lung mechanics, with potential implications for lung bioengineering.

Keywords: Gamma irradiation, Lung bioengineering, Lung decellularization, Organ scaffold, Pulmonary mechanics, Decellularization, Gamma irradiation, Mouse lung, Pulmonary mechanics, dodecyl sulfate sodium, animal tissue, Article, artificial ventilation, bioengineering, bioreactor, compliance (physical), controlled study, freezing, gamma irradiation, lung, lung mechanics, lung resistance, male, mouse, nonhuman, room temperature, scanning electron microscopy, tissue scaffold, trachea pressure

Le Roux, D., Burger, P. B., Niemand, J., Grobler, A., Urbán, P., Fernàndez-Busquets, X., Barker, R. H., Serrano, A. E., I. Louw, A., Birkholtz, L. M., (2014). Novel S-adenosyl-L-methionine decarboxylase inhibitors as potent antiproliferative agents against intraerythrocytic Plasmodium falciparum parasites International Journal for Parasitology: Drugs and Drug Resistance , 4, (1), 28-36

S-adenosyl-l-methionine decarboxylase (AdoMetDC) in the polyamine biosynthesis pathway has been identified as a suitable drug target in Plasmodium falciparum parasites, which causes the most lethal form of malaria. Derivatives of an irreversible inhibitor of this enzyme, 5'-{[(Z)-4-amino-2-butenyl]methylamino}-5'-deoxyadenosine (MDL73811), have been developed with improved pharmacokinetic profiles and activity against related parasites, Trypanosoma brucei. Here, these derivatives were assayed for inhibition of AdoMetDC from P. falciparum parasites and the methylated derivative, 8-methyl-5'-{[(Z)-4-aminobut-2-enyl]methylamino}-5'-deoxyadenosine (Genz-644131) was shown to be the most active. The in vitro efficacy of Genz-644131 was markedly increased by nanoencapsulation in immunoliposomes, which specifically targeted intraerythrocytic P. falciparum parasites.

Keywords: Immunoliposomes, Plasmodium, Polyamines, S-adenosyl-l-methionine decarboxylase

Sachot, N., Engel, E., Castaño, O., (2014). Hybrid organic-inorganic scaffolding biomaterials for regenerative therapies Current Organic Chemistry , 18, (18), 2299-2314

The introduction of hybrid materials in regenerative medicine has solved some problems related to the mechanical and bioactive properties of biomaterials. Calcium phosphates and their derivatives have provided the basis for inorganic components, thanks to their good bioactivity, especially in bone regeneration. When mixed with biodegradable polymers, the result is a synergic association that mimics the composition of many tissues of the human body and, additionally, exhibits suitable mechanical properties. Together with the development of nanotechnology and new synthesis methods, hybrids offer a promising option for the development of a third or fourth generation of smart biomaterials and scaffolds to guide the regeneration of natural tissues, with an optimum efficiency/cost ratio. Their potential bioactivity, as well as other valuable features of hybrids, open promising new pathways for their use in bone regeneration and other tissue repair therapies. This review provides a comprehensive overview of the different hybrid organic-inorganic scaffolding biomaterials developed so far for regenerative therapies, especially in bone. It also looks at the potential for research into hybrid materials for other, softer tissues, which is still at an initial stage, but with very promising results.

Keywords: Biodegradable polymer, Hybrid materials, Nanoparticles, Ormoglass

Álvarez, Z., Sena, E., Mattotti, M., Engel, E., Alcántara, S., (2014). An efficient and reproducible method to culture Bergmann and cortical radial glia using textured PMMA Journal of Neuroscience Methods , 232, 93-101

Background: Radial glia cells comprise the principal population of neural stem cells (NSC) during development. Attempts to develop reproducible radial glia and NSC culture methods have met with variable results, yielding non-adherent cultures or requiring the addition of growth factors. Recent studies demonstrated that a 2-μm patterned poly-methyl methacrylate (ln2 PMMA) grooved scaffold, by mimicking the biophysical and microtopographic properties of the embryonic NSC niche, induces the de-differentiation of glial cells into functional radial glia cells. New method: Here we describe a method for obtaining cultures of adherent Bergmann radial glia (BRG) and cortical radial glia (CRG). The growth substrate is ln2 PMMA and the addition of growth factors is not required. Results: Postnatal glia obtained from mouse cerebellum or cerebral cortex and grown on ln2 PMMA adopted a BRG/CRG phenotype characterized by a bipolar shape, the up-regulation of progenitor markers such as nestin and Sox2, and the down-regulation of vimentin and GFAP. Neurons cultured over the BRG/CRG aligned their processes with those of the glial shafts, thus mimicking the behavior of migrating neuronal cells. Comparison with existing methods: The ln2 PMMA culture method offers an ideal system for analyzing both the biochemical factors controlling the neurogenic potential of BRG/CRG and neuronal migration. Conclusions: The ln2 PMMA method is a reproducible system to obtain immature BRG/CRG preparations in vitro. It can be used to study the properties of CNS progenitor cells as well as the interactions between radial glia and neurons, and supports cultured progenitors for use in different applications. © 2014 Elsevier B.V.

Keywords: Astrocytes, Bergmann glia, Micro-patterning, Poly-methyl methacrylate (PMMA), Progenitors, Radial glia, Surface topography

Sánchez Egea, Antonio J., Valera, Marius, Parraga Quiroga, Juan Manuel, Proubasta, Ignasi, Noailly, J., Lacroix, Damien, (2014). Impact of hip anatomical variations on the cartilage stress: A finite element analysis towards the biomechanical exploration of the factors that may explain primary hip arthritis in morphologically normal subjects Clinical Biomechanics , 29, (4), 444-450

AbstractBackground Hip arthritis is a pathology linked to hip-cartilage degeneration. Although the aetiology of this disease is not well defined, it is known that age is a determinant risk factor. However, hip arthritis in young patients could be largely promoted by biomechanical factors. The objective of this paper is to analyze the impact of some normal anatomical variations on the cartilage stress distributions numerically predicted at the hip joint during walking. Methods A three-dimensional finite element model of the femur and the pelvis with the most relevant axial components of muscle forces was used to simulate normal walking activity. The hip anatomical condition was defined by: neck shaft angle, femoral anteversion angle, and acetabular anteversion angle with a range of 110-130º, 0-20º, and 0-20º, respectively. The direct boundary method was used to simulate the hip contact. Findings The hydrostatic stress found at the cartilage and labrum showed that a ± 10º variation with respect to the reference brings significant differences between the anatomic models. Acetabular anteversion angle of 0º and femoral anteversion angle of 0º were the most affected anatomical conditions with values of hydrostatic stress in the cartilage near 5 MPa under compression. Interpretation Cartilage stresses and contact areas were equivalent to the results found in literature and the most critical anatomical regions in terms of tissue loads were in a good accordance with clinical evidence. Altogether, results showed that decreasing femoral or acetabular anteversion angles isolately causes a dramatic increase in cartilage loads.

Keywords: Hip arthritis, Neck shaft angle, Femoral and acetabular anteversions, Cartilage load, Hip joint contact, Finite element analysis

Oller-Moreno, S., Pardo, A., Jimenez-Soto, J. M., Samitier, J., Marco, S., (2014). Adaptive Asymmetric Least Squares baseline estimation for analytical instruments SSD 2014 Proceedings 11th International Multi-Conference on Systems, Signals & Devices (SSD) , IEEE (Castelldefels-Barcelona, Spain) , 1569846703

Automated signal processing in analytical instrumentation is today required for the analysis of highly complex biomedical samples. Baseline estimation techniques are often used to correct long term instrument contamination or degradation. They are essential for accurate peak area integration. Some methods approach the baseline estimation iteratively, trying to ignore peaks which do not belong to the baseline. The proposed method in this work consists of a modification of the Asymmetric Least Squares (ALS) baseline removal technique developed by Eilers and Boelens. The ALS technique suffers from bias in the presence of intense peaks (in relation to the noise level). This is typical of diverse instrumental techniques such as Gas Chromatography-Mass Spectrometry (GC-MS) or Gas Chromatography-Ion Mobility Spectrometry (GC-IMS). In this work, we propose a modification (named psalsa) to the asymmetry weights of the original ALS method in order to better reject large peaks above the baseline. Our method will be compared to several versions of the ALS algorithm using synthetic and real GC signals. Results show that our proposal improves previous versions being more robust to parameter variations and providing more accurate peak areas.

Keywords: Gas chromatography, Instruments, Radioactivity measurement, Signal processing, Analytical instrument, Analytical Instrumentation, Asymmetric least squares, Baseline estimation, Baseline removal, Gas chromatography-mass spectrometries (GC-MS), Instrumental techniques, Noise levels, Iterative methods

Lozano-Garcia, M., Fiz, J. A., Jané, R., (2014). Analysis of normal and continuous adventitious sounds for the assessment of asthma IFMBE Proceedings XIII Mediterranean Conference on Medical and Biological Engineering and Computing 2013 (ed. Roa Romero, Laura M.), Springer International Publishing (London, UK) 41, 981-984

Assessment of asthma is a difficult procedure which is based on the correlation of multiple factors. A major component in the diagnosis of asthma is the assessment of BD response, which is performed by traditional spirometry. In this context, the analysis of respiratory sounds (RS) provides relevant and complementary information about the function of the respiratory system. In particular, continuous adventitious sounds (CAS), such as wheezes, contribute to assess the severity of patients with obstructive diseases. On the other hand, the intensity of normal RS is dependent on airflow level and, therefore, it changes depending on the level of obstruction. This study proposes a new approach to RS analysis for the assessment of asthmatic patients, by combining the quantification of CAS and the analysis of the changes in the normal sound intensity-airflow relationship. According to results obtained from three patients with different characteristics, the proposed technique seems more sensitive and promising for the assessment of asthma.

Keywords: Asthma, Bronchodilator response, Continuous adventitious sound, Respiratory sound intensity, Wheezes

Torres, A., Fiz, J. A., Jané, R., (2014). Cancellation of cardiac interference in diaphragm EMG signals using an estimate of ECG reference signal IFMBE Proceedings XIII Mediterranean Conference on Medical and Biological Engineering and Computing 2013 (ed. Roa Romero, Laura M.), Springer International Publishing (London, UK) 41, 1000-1004

The analysis of the electromyographic signal of the diaphragm muscle (EMGdi) can provide important information in order to evaluate the respiratory muscular function. However, EMGdi signals are usually contaminated by the electrocardiographic (ECG) signal. An adaptive noise cancellation (ANC) based on event-synchronous cancellation can be used to reduce the ECG interference in the recorded EMGdi activity. In this paper, it is proposed an ANC scheme for cancelling the ECG interference in EMGdi signals using only the EMGdi signal (without acquiring the ECG signal). In this case the detection of the QRS complex has been performed directly in the EMGdi signal, and the ANC algorithm must be robust to false or missing QRS detections. Furthermore, an automatic criterion to select the adaptive constant of the LMS algorithm has been proposed (μ). The μ constant is selected automatically so that the canceling signal energy equals the energy of the reference signal (which is an estimation of the ECG interference present in the EMGdi signal). This approach optimizes the tradeoff between cancellation of ECG interference and attenuation of EMG component. A number of weights equivalent of a time window that contains several QRS complexes is selected in order to make the algorithm robust to QRS detection errors.

Keywords: Adaptive Canceller, EMG, Diaphragm muscle

Aviles, A. I., Casals, A., (2014). Interpolation based deformation model for minimally invasive beating heart surgery IFMBE Proceedings XIII Mediterranean Conference on Medical and Biological Engineering and Computing 2013 (ed. Roa Romero, Laura M.), Springer International Publishing (London, UK) 41, 372-375

Heart motion compensation is a key issue in medical robotics due to the benefits that minimally invasive beating heart surgery offers over traditional cardiac surgery. Although different proposals have been presented, nowadays, there is not yet a suitable solution working in real clinical environments due to the lack of robustness of existing methods. The process of heart motion estimation required to produce the compensation actions can be tackled as a process of three iterative steps. The first based on generating a deformation model from the processing of a video sequence of the beating heart. The selection of a deformation model is crucial in the sense that it has to offer both valuable information and good computational performance. These characteristics are required when the reaction time has a significant repercussion over the system behavior, as in this case. This paper, presents a computational analysis of deformation model based on interpolation methods. In particular, wavelet and thin-plate splines are evaluated. The significance of this study relies on the fact that it is a reference starting point of reference for creating both a common framework and a robust solution. In addition, the obtained results will contribute to increase the robustness from the initial stage of the solution.

Keywords: Deformation model, Wavelets, Computer performance, Radial basis functions, Interpolation methods

Álvarez, Zaida, Mateos-Timoneda, Miguel A., Hyrossová, Petra, Castaño, Oscar, Planell, Josep A., Perales, José C., Engel, Elisabeth, Alcántara, Soledad, (2013). The effect of the composition of PLA films and lactate release on glial and neuronal maturation and the maintenance of the neuronal progenitor niche Biomaterials 34, (9), 2221-2233

To develop tissue engineering strategies useful for repairing damage in the central nervous system (CNS) it is essential to design scaffolds that emulate the NSC niche and its tight control of neural cell genesis, growth, and differentiation. In this study we tested two types of poly l/dl lactic acid (PLA95/5 and PLA70/30), a biodegradable material permissive for neural cell adhesion and growth, as materials for nerve regeneration. Both PLA were slightly hydrophobic and negatively charged but differed in crystallinity, stiffness and degradation rate. PLA95/5 films were highly crystalline, stiff (GPa), and did not degrade significantly in the one-month period analyzed in culture. In contrast, PLA70/30 films were more amorphous, softer (MPa) and degraded faster, releasing significant amounts of lactate into the culture medium. PLA70/30 performs better than PLA95/5 for primary cortical neural cell adhesion, proliferation and differentiation, maintaining the pools of neuronal and glial progenitor cells in vitro. l-lactate in the medium recapitulated PLA70/30's maintenance of neuronal restricted progenitors but did not sustain bipotential or glial restricted progenitors in the cultures, as occurred when neural cells were grown on PLA70/30. Our results suggest that PLA70/30 may mimic some of the physical and biochemical characteristics of the NSC niche. Its mechanical and surface properties may act synergistically in the modulation of bipotential and glial restricted progenitor phenotypes, while it is l-lactate, either added to the medium or released by the film that drives the maintenance of neuronal restricted progenitor cell phenotypes.

Keywords: Polylactic acid, Degradation, Neurons, Progenitors, Lactate, Glial cells, NSC niche

Serra, T., Planell, J. A., Navarro, M., (2013). High-resolution PLA-based composite scaffolds via 3-D printing technology Acta Biomaterialia 9, (3), 5521-5530

Fabrication of new biodegradable scaffolds that guide and stimulate tissue regeneration is still a major issue in tissue engineering approaches. Scaffolds that possess adequate biodegradability, pore size, interconnectivity, bioactivity and mechanical properties in accordance with the injured tissue are required. This work aimed to develop and characterize three-dimensional (3-D) scaffolds that fulfill the aforementioned requirements. For this, a nozzle-based rapid prototyping system was used to combine polylactic acid and a bioactive CaP glass to fabricate 3-D biodegradable scaffolds with two patterns (orthogonal and displaced double layer). Scanning electron microscopy and micro-computer tomography showed that 3-D scaffolds had completely interconnected porosity, uniform distribution of the glass particles, and a controlled and repetitive architecture. Surface properties were also assessed, showing that the incorporation of glass particles increased both the roughness and the hydrophilicity of the scaffolds. Mechanical tests indicated that compression strength is dependent on the scaffold geometry and the presence of glass. Preliminary cell response was studied with primary mesenchymal stem cells (MSC) and revealed that CaP glass improved cell adhesion. Overall, the results showed the suitability of the technique/materials combination to develop 3-D porous scaffolds and their initial biocompatibility, both being valuable characteristics for tissue engineering applications.

Keywords: Rapid prototyping, Scaffold, Polylactic acid, Biodegradable, Composite

Ruiz, C., Noailly, J., Lacroix, D., (2013). Material property discontinuities in intervertebral disc porohyperelastic finite element models generate numerical instabilities due to volumetric strain variations Journal of the Mechanical Behavior of Biomedical Materials , 26, 1-10

Numerical studies of the intervertebral disc (IVD) are important to better understand the load transfer and the mechanobiological processes within the disc. Among the relevant calculations, fluid-related outputs are critical to describe and explore accurately the tissue properties. Porohyperelastic finite element models of IVD can describe accurately the disc behaviour at the organ level and allow the inclusion of fluid effects. However, results may be affected by numerical instabilities when fast load rates are applied. We hypothesized that such instabilities would appear preferentially at material discontinuities such as the annulus-nucleus boundary and should be considered when testing mesh convergence. A L4-L5 IVD model including the nucleus, annulus and cartilage endplates were tested under pure rotational loads, with different levels of mesh refinement. The effect of load relaxation and swelling were also studied. Simulations indicated that fluid velocity oscillations appeared due to numerical instability of the pore pressure spatial derivative at material discontinuities. Applying local refinement only was not enough to eliminate these oscillations. In fact, mesh refinements had to be local, material-dependent, and supplemented by the creation of a material transition zone, including interpolated material properties. Results also indicated that oscillations vanished along load relaxation, and faster attenuation occurred with the incorporation of the osmotic pressure. We concluded that material discontinuities are a major cause of instability for poromechanical calculations in multi-tissue models when load velocities are simulated. A strategy was presented to address these instabilities and recommendations on the use of IVD porohyperelastic models were given.

Keywords: Fast loads, Intervertebral disc, Numerical instabilities, Poroelastic model

Rodríguez-Hernández, Ana G., Muñoz-Tabares, José, Godoy-Gallardo, Maria, Juárez, Antonio, Gil, Francisco-Javier, (2013). S. sanguinis adhesion on rough titanium surfaces: Effect of culture media Materials Science and Engineering: C 33, (2), 714-720

Bacterial colonization plays a key role in dental implant failure, because they attach directly on implant surface upon implantation. Between different types of bacteria associated with the oral environment, Streptococcus sanguinis is essential in this process since it is an early colonizer. In this work the relationship between titanium surfaces modified by shot blasting treatment and S. sanguinis adhesion; have been studied in approached human mouth environment. Bacteria pre-inoculated with routinary solution were put in contact with titanium samples, shot-blasted with alumina and silicon carbide, and adhesion results were compared with those obtained when bacteria were pre-inoculated with modified artificial saliva medium and on saliva pre-coated titanium samples. Our results showed that bacterial adhesion on titanium samples was influenced by culture conditions. When S. sanguinis was inoculated in routinary culture media, colonies forming unities per square millimeter presented an increment correlated with roughness and surface energy, but separated by the type of particle used during shot-blasting treatment; whereas in modified artificial saliva only a relationship between bacteria adhered and the increment in both roughness and surface energy were observed, regardless of the particle type. Finally, on human saliva pre-coated samples no significant differences were observed among roughness, surface energy or particle.

Keywords: S. sanguinis, Bacterial adhesion, Titanium, Artificial saliva, Surface energy, Roughness

Peñuelas, O., Melo, E., Sánchez, C., Sánchez, I., Quinn, K., Ferruelo, A., Pérez-Vizcaíno, F., Esteban, A., Navajas, D., Nin, N., Lorente, J. A., Farré, R., (2013). Antioxidant effect of human adult adipose-derived stromal stem cells in alveolar epithelial cells undergoing stretch Respiratory Physiology & Neurobiology , 188, (1), 1-8

Introduction: Alveolar epithelial cells undergo stretching during mechanical ventilation. Stretch can modify the oxidative balance in the alveolar epithelium. The aim of the present study was to evaluate the antioxidant role of human adult adipose tissue-derived stromal cells (hADSCs) when human alveolar epithelial cells were subjected to injurious cyclic overstretching. Methods: A549 cells were subjected to biaxial stretch (0-15% change in surface area for 24. h, 0.2. Hz) with and without hADSCs. At the end of the experiments, oxidative stress was measured as superoxide generation using positive nuclear dihydroethidium (DHE) staining, superoxide dismutase (SOD) activity in cell lysates, 8-isoprostane concentrations in supernatant, and 3-nitrotyrosine by indirect immunofluorescence in fixed cells. Results: Cyclically stretching of AECs induced a significant decrease in SOD activity, and an increase in 8-isoprostane concentrations, DHE staining and 3-nitrotyrosine staining compared with non-stretched cells. Treatment with hADSCs significantly attenuated stretch-induced changes in SOD activity, 8-isoprostane concentrations, DHE and 3-nitrotyrosine staining. Conclusion: These data suggest that hADSCs have an anti-oxidative effect in human alveolar epithelial cells undergoing cyclic stretch.

Keywords: Acute lung injury, Cyclic stretch, Human adipose-derived stromal stem cells, Oxidative stress

Almendros, Isaac, Carreras, Alba, Montserrat, Josep M., Gozal, David, Navajas, Daniel, Farre, Ramon, (2012). Potential role of adult stem cells in obstructive sleep apnea Frontiers in Neurology 3, 1-6

Adult stem cells are undifferentiated cells that can be mobilized from the bone marrow or other organs, home into injured tissues and differentiate into different cell phenotypes to serve in a repairing capacity. Furthermore, these cells can respond to inflammation and oxidative stress by exhibiting immunomodulatory properties. The protective and reparative roles of mesenchymal stem cells (MSCs), very small embryonic-like stem cells (VSELs) and endothelial progenitor cells (EPCs) have primarily been examined and characterized in auto-immune and cardiovascular diseases. Obstructive sleep apnea (OSA) is a very prevalent disease (4-5% of adult population and 2-3% of children) characterized by an abnormal increase in upper airway collapsibility. Recurrent airway obstructions elicit arterial oxygen desaturations, increased inspiratory efforts and sleep fragmentation, which have been associated with important long-term neurocognitive, metabolic, and cardiovascular consequences. Since inflammation, oxidative stress and endothelial dysfunction are key factors in the development of the morbid consequences of OSA, bone marrow-derived stem cells could be important modulators of the morbid phenotype by affording a protective role. This mini-review is focused on the recent data available on EPCs, VSELs and MSCs in both animal models and patients with OSA.

Keywords: Mesenchymal Stem Cells, Sleep Apnea, Endothelial progenitor cells, Very Small-like Embryonic Stem Cells, Adult bone-marrow derived stem cells

Bakker, G. J., Eich, C., Torreno-Pina, J. A., Diez-Ahedo, R., Perez-Samper, G., Van Zanten, T. S., Figdor, C. G., Cambi, A., Garcia-Parajo, M. F., (2012). Lateral mobility of individual integrin nanoclusters orchestrates the onset for leukocyte adhesion Proceedings of the National Academy of Sciences of the United States of America 109, (13), 4869-4874

Integrins are cell membrane adhesion receptors involved in morphogenesis, immunity, tissue healing, and metastasis. A central, yet unresolved question regarding the function of integrins is how these receptors regulate both their conformation and dynamic nanoscale organization on the membrane to generate adhesion-competent microclusters upon ligand binding. Here we exploit the high spatial (nanometer) accuracy and temporal resolution of single-dye tracking to dissect the relationship between conformational state, lateral mobility, and microclustering of the integrin receptor lymphocyte function-associated antigen 1 (LFA-1) expressed on immune cells. We recently showed that in quiescent monocytes, LFA-1 preorganizes in nanoclusters proximal to nanoscale raft components. We now show that these nanoclusters are primarily mobile on the cell surface with a small (ca. 5%) subset of conformational- active LFA-1 nanoclusters preanchored to the cytoskeleton. Lateral mobility resulted crucial for the formation of microclusters upon ligand binding and for stable adhesion under shear flow. Activation of high-affinity LFA-1 by extracellular Ca 2+ resulted in an eightfold increase on the percentage of immobile nanoclusters and cytoskeleton anchorage. Although having the ability to bind to their ligands, these active nanoclusters failed to support firm adhesion in static and low shear-flow conditions because mobility and clustering capacity were highly compromised. Altogether, our work demonstrates an intricate coupling between conformation and lateral diffusion of LFA-1 and further underscores the crucial role of mobility for the onset of LFA-1 mediated leukocyte adhesion.

Keywords: Cumulative probability distribution, Integrin lymphocyte function-associated antigen 1, Intercellular adhesion molecule, Single molecule detection

Lagunas, Anna , Comelles, Jordi, Martínez, Elena, Prats-Alfonso, Elisabet , Acosta, Gerardo A., Albericio, Fernando , Samitier, Josep , (2012). Cell adhesion and focal contact formation on linear RGD molecular gradients: study of non-linear concentration dependence effects Nanomedicine: Nanotechnology, Biology and Medicine , 8, (4), 432-439

Cell adhesion onto bioengineered surfaces is affected by a number of variables, including the former substrate derivatization process. In this investigation, we studied the correlation between cell adhesion and cell–adhesive ligand surface concentration and organization due to substrate modification. For this purpose, Arg-Gly-Asp (RGD) gradient surfaces were created on poly(methyl methacrylate) substrates by continuous hydrolysis and were then grafted with biotin-PEG-RGD molecules. Cell culture showed that adhesion behavior changes in a nonlinear way in the narrow range of RGD surface densities assayed (2.8 to 4.4 pmol/cm2), with a threshold value of 4.0 pmol/cm2 for successful cell attachment and spreading. This nonlinear dependence may be explained by nonhomogeneous RGD surface distribution at the nanometre scale, conditioned by the stochastic nature of the hydrolysis process. Atomic force microscopy analysis of the gradient surface showed an evolution of surface morphology compatible with this hypothesis.

Keywords: RGD gradient, Cell adhesion, Poly(methyl methacrylate), Hydrolysis, Biotin-streptavidin

Roca-Cusachs, P., Iskratsch, T., Sheetz, M. P., (2012). Finding the weakest link: exploring integrin-mediated mechanical molecular pathways Journal of Cell Science 125, (13), 3025-3038

From the extracellular matrix to the cytoskeleton, a network of molecular links connects cells to their environment. Molecules in this network transmit and detect mechanical forces, which subsequently determine cell behavior and fate. Here, we reconstruct the mechanical pathway followed by these forces. From matrix proteins to actin through integrins and adaptor proteins, we review how forces affect the lifetime of bonds and stretch or alter the conformation of proteins, and how these mechanical changes are converted into biochemical signals in mechanotransduction events. We evaluate which of the proteins in the network can participate in mechanotransduction and which are simply responsible for transmitting forces in a dynamic network. Besides their individual properties, we also analyze how the mechanical responses of a protein are determined by their serial connections from the matrix to actin, their parallel connections in integrin clusters and by the rate at which force is applied to them. All these define mechanical molecular pathways in cells, which are emerging as key regulators of cell function alongside better studied biochemical pathways.

Keywords: Cell adhesion, Cytoskeleton, Mechanotransduction

Dries, Koen, Helden, Suzanne, Riet, Joostte, Diez-Ahedo, Ruth, Manzo, Carlo, Oud, Machteld, Leeuwen, Frank, Brock, Roland, Garcia-Parajo, Maria, Cambi, Alessandra, Figdor, CarlG, (2012). Geometry sensing by dendritic cells dictates spatial organization and PGE2-induced dissolution of podosomes Cellular and Molecular Life Sciences , 69, (11), 1889-1901

Assembly and disassembly of adhesion structures such as focal adhesions (FAs) and podosomes regulate cell adhesion and differentiation. On antigen-presenting dendritic cells (DCs), acquisition of a migratory and immunostimulatory phenotype depends on podosome dissolution by prostaglandin E2 (PGE2). Whereas the effects of physico-chemical and topographical cues have been extensively studied on FAs, little is known about how podosomes respond to these signals. Here, we show that, unlike for FAs, podosome formation is not controlled by substrate physico-chemical properties. We demonstrate that cell adhesion is the only prerequisite for podosome formation and that substrate availability dictates podosome density. Interestingly, we show that DCs sense 3-dimensional (3-D) geometry by aligning podosomes along the edges of 3-D micropatterned surfaces. Finally, whereas on a 2-dimensional (2-D) surface PGE2 causes a rapid increase in activated RhoA levels leading to fast podosome dissolution, 3-D geometric cues prevent PGE2-mediated RhoA activation resulting in impaired podosome dissolution even after prolonged stimulation. Our findings indicate that 2-D and 3-D geometric cues control the spatial organization of podosomes. More importantly, our studies demonstrate the importance of substrate dimensionality in regulating podosome dissolution and suggest that substrate dimensionality plays an important role in controlling DC activation, a key process in initiating immune responses.

Keywords: Mechanosensitivity, Podosomes, Dendritic cell, Adhesion

Yang, Cheng, Lates, Vasilica, Prieto-Simón, Beatriz, Marty, Jean-Louis, Yang, Xiurong, (2012). Aptamer-DNAzyme hairpins for biosensing of Ochratoxin A Biosensors and Bioelectronics 32, (1), 208-212

We report an aptasensor for biosensing of Ochratoxin A (OTA) using aptamer-DNAzyme hairpin as biorecognition element. The structure of this engineered nucleic acid includes the horseradish peroxidase (HRP)-mimicking DNAzyme and the OTA specific aptamer sequences. A blocking tail captures a part of these sequences in the stem region of the hairpin. In the presence of OTA, the hairpin is opened due to the formation of the aptamer–analyte complex. As a result, self-assembly of the active HRP-mimicking DNAzyme occurs. The activity of this DNAzyme is linearly correlated with OTA concentration up to 10 nM, showing a limit of detection of 2.5 nM.

Keywords: Ochratoxin A, Aptamer, G-quadruplex, DNAzyme, Hairpin

Levato, Riccardo, Mateos-Timoneda, Miguel A., Planell, Josep A., (2012). Preparation of biodegradable polylactide microparticles via a biocompatible procedure Macromolecular Bioscience 12, (4), 557-566

PLA MPs are prepared via a novel and toxic-chemical-free fabrication route using ethyl lactate, a green solvent and FDA-approved aroma. MPs are obtained by a solution jet break-up and solvent displacement method. Adjusting flow parameters allows the tuning of MPs size between 60 and 180 µm, with reduced polydispersity. Morphological analysis shows microporous particles with Janus-like surface. A fluorophore is successfully loaded into the MPs during their formation step. This versatile green solvent-based procedure is proven to be suitable for drug encapsulation and delivery applications. The method may be extended to different droplet generation techniques.

Keywords: Biocompatibility, Biodegradable, Green solvents, Microparticles, Poly(lactic acid)

Juanola-Feliu, E., Colomer-Farrarons, J., Miribel-Català , P., Samitier, J., Valls-Pasola, J., (2012). Market challenges facing academic research in commercializing nano-enabled implantable devices for in-vivo biomedical analysis Technovation , 32, (3-4), 193-204

This article reports on the research and development of a cutting-edge biomedical device for continuous in-vivo glucose monitoring. This entirely public-funded process of technological innovation has been conducted at the University of Barcelona within a context of converging technologies involving the fields of medicine, physics, chemistry, biology, telecommunications, electronics and energy. The authors examine the value chain and the market challenges faced by in-vivo implantable biomedical devices based on nanotechnologies. In so doing, they trace the process from the point of applied research to the final integration and commercialization of the product, when the social rate of return from academic research can be estimated. Using a case-study approach, the paper also examines the high-tech activities involved in the development of this nano-enabled device and describes the technology and innovation management process within the value chain conducted in a University-Hospital-Industry-Administration-Citizens framework. Here, nanotechnology is seen to represent a new industrial revolution, boosting the biomedical devices market. Nanosensors may well provide the tools required for investigating biological processes at the cellular level in vivo when embedded into medical devices of small dimensions, using biocompatible materials, and requiring reliable and targeted biosensors, high speed data transfer, safely stored data, and even energy autonomy.

Keywords: Biomedical device, Diabetes, Innovation management, Nanobiosensor, Nanotechnology, Research commercialization, Technology transfer, Academic research, Applied research, Barcelona, Biocompatible materials, Biological process, Biomedical analysis, Biomedical devices, Cellular levels, Converging technologies, Glucose monitoring, High-speed data transfer, Implantable biomedical devices, Implantable devices, In-vivo, Industrial revolutions, Innovation management, Medical Devices, Nanobiosensor, Rate of return, Research and development, Technological innovation, Value chains, Biological materials, Biomedical engineering, Biosensors, Commerce, Data transfer, Earnings, Engineering education, Glucose, Implants (surgical), Industrial research, Innovation, Medical problems, Nanosensors, Nanotechnology, Technology transfer, Equipment

Pegueroles, M., Tonda-Turo, C., Planell, J. A., Gil, F. J., Aparicio, C., (2012). Adsorption of fibronectin, fibrinogen, and albumin on TiO2: Time-resolved kinetics, structural changes, and competition study Biointerphases , 7, (48), 13

An understanding of protein adsorption process is crucial for designing biomaterial surfaces. In this work, with the use of a quartz-crystal microbalance with dissipation monitoring, we researched the following: (a) the kinetics of adsorption on TiO2 surfaces of three extensively described proteins that are relevant for metallic implant integration [i.e., albumin (BSA), fibrinogen (Fbg), and fibronectin (Fn)]; and (b) the competition of those proteins for adsorbing on TiO2 in a two-step experiment consisted of sequentially exposing the surfaces to different monoprotein solutions. Each protein showed a different process of adsorption and properties of the adlayer-calculated using the Voigt model. The competition experiments showed that BSA displaced larger proteins such as Fn and Fbg when BSA was introduced as the second protein in the system, whereas the larger proteins laid on top of BSA forming an adsorbed protein bi-layer when those were introduced secondly in the system.

Keywords: QCM, Human plasma fibronectin, Induced conformational-changes, Von-willebrand-factor, BSA, Protein adsortion, Polymer surfaces, Solid-surfaces, Viscoelastic properties, Globular-proteins

Fazel Zarandi, M. H., Avazbeigi, M., (2012). A multi-agent solution for reduction of bullwhip effect in fuzzy supply chains Journal of Intelligent and Fuzzy Systems , 23, (5), 259-268

In this paper, we present a new Multi-Agent System for reduction of the bullwhip effect in fuzzy supply chains. First, we show that a supply chain that uses an optimal ordering policy without data sharing among echelons still suffers from the bullwhip effect. Then, we propose the multi-agent solution to manage and reduce the bullwhip effect. The proposed multi-agent system includes four different types of agents in which each agent has its own list of actions. The proposed Multi-agent System applies a new Tabu Search algorithm for fuzzy rule generation, and a new data filtering algorithm for extraction of the bullwhip-free data from supply chain data warehouse. We validate the multi-agent system under different conditions and discuss how the system responds to different factors. The results show that the proposed multi-agent system reduces the bullwhip effect significantly in a rational time.

Keywords: Bullwhip effect, Bullwhip-free data, Decentralized decision making, Fuzzy rule base, Fuzzy supply chain, Fuzzy time series, Multi-agent system, Supply chain management

Sarlabous, L., Torres, A., Fiz, J. A., Morera, J., Jané, R., (2012). Evaluation and adaptive attenuation of the cardiac vibration interference in mechanomyographic signals Engineering in Medicine and Biology Society (EMBC) 34th Annual International Conference of the IEEE , IEEE (San Diego, USA) , 3400-3403

The study of the mechanomyographic signal of the diaphragm muscle (MMGdi) is a promising technique in order to evaluate the respiratory muscles effort. The relationship between amplitude and frequency parameters of this signal with the respiratory effort performed during respiration is of great interest for researchers and physicians due to its diagnostic potentials. However, MMGdi signals are frequently contaminated by a cardiac vibration or mechanocardiographic (MCG) signal. An adaptive noise cancellation (ANC) can be used to reduce the MCG interference in the recorded MMGdi activity. In this paper, it is evaluated the proposed ANC scheme by means of a synthetic MMGdi signal with a controlled MCG interference. The Pearson's correlation coefficient (PCC) between both root mean square (RMS) and mean frequency (fm) of the synthetic MMGdi signal are considerably reduced with the presence of cardiac vibration noise (from 0.95 to 0.87, and from 0.97 to 0.76, respectively). With the ANC algorithm proposed the effect of the MCG noise on the amplitude and frequency of MMG parameters is reduced considerably (PCC of 0.93 and 0.97 for the RMS and fm, respectively). The ANC method proposed in this work is an interesting technique to attenuate the cardiac interference in respiratory MMG signals. Further investigation should be carried out to evaluate the performance of the ANC algorithm in real MMGdi signals.

Keywords: Adaptive filters, Frequency modulation, Interference, Muscles, Noise cancellation, Vibrations, Cardiology, Medical signal processing, Muscle, Signal denoising, ANC algorithm, MCG interference, Pearson correlation coefficient, Adaptive noise cancellation, Cardiac vibration interference, Cardiac vibration noise, Diaphragm muscle, Mechanocardiographic signal, Mechanomyographic signals, Respiratory muscles effort

Serra, T., Navarro, M., Planell, J. A., (2012). Fabrication and characterization of biodegradable composite scaffolds for tissue engineering Innovative Developments in Virtual and Physical Prototyping 5th International Conference on Advanced Research and Rapid Prototyping (ed. Margarida, T., Ferreira, D.), Taylor & Francis (Leiria, Portugal) VR@P, 67-72

In this study, polylactic acid (PLA) and polyethylene glycol (PEG) were combined with soluble CaP glass particles and processed by rapid prototyping to obtain fully biodegradable structures for Tissue Engineering applications. The obtained 3D biodegradable structures were characterized in terms of their architecture and mechanical properties. The scaffold morphology, internal micro-architecture and mechanical properties were evaluated using Scanning Electron Microscopy (SEM), micro-computed tomography (micro-CT) and mechanical testing, respectively. Well defined structures with pore size of 350-400μm (in the axial view), struts width of approximately 70-80μm, and a porosity ranging between 60-65% were obtained. The combination RP and PLA/PEG/CaP glass turned into promising fully degradable, mechanically stable, bioactive and biocompatible composite scaffolds for TE.

Keywords: Axial view, Biodegradable composites, Composite scaffolds, Glass particles, Mechanically stable, Micro architectures, Micro computed tomography (micro-CT), Poly lactic acid, Scaffold morphology, Tissue engineering applications, Well-defined structures, Bioactive glass, Mechanical properties, Mechanical testing, Polyethylene glycols, Polymer blends, Rapid prototyping, Scaffolds (biology), Scanning electron microscopy, Computerized tomography

Mesquita, J., Poree, F., Carrault, G., Fiz, J. A., Abad, J., Jané, R., (2012). Respiratory and spontaneous arousals in patients with Sleep Apnea Hypopnea Syndrome Engineering in Medicine and Biology Society (EMBC) 34th Annual International Conference of the IEEE , IEEE (San Diego, USA) , 6337-6340

Sleep in patients with Sleep Apnea-Hypopnea Syndrome (SAHS) is frequently interrupted with arousals. Increased amounts of arousals result in shortening total sleep time and repeated sleep-arousal change can result in sleep fragmentation. According to the American Sleep Disorders Association (ASDA) an arousal is a marker of sleep disruption representing a detrimental and harmful feature for sleep. The nature of arousals and its role on the regulation of the sleep process raises controversy and has sparked the debate in the last years. In this work, we analyzed and compared the EEG spectral content of respiratory and spontaneous arousals on a database of 45 SAHS subjects. A total of 3980 arousals (1996 respiratory and 1984 spontaneous) were analyzed. The results showed no differences between the spectral content of the two kinds of arousals. Our findings raise doubt as to whether these two kinds of arousals are truly triggered by different organic mechanisms. Furthermore, they may also challenge the current beliefs regarding the underestimation of the importance of spontaneous arousals and their contribution to sleep fragmentation in patients suffering from SAHS.

Keywords: Adaptive filters, Correlation, Databases, Electroencephalography, Hospitals, Sleep apnea, Electroencephalography, Medical signal processing, Pneumodynamics, Sleep, EEG spectral content, Organic mechanism, Respiratory, Sleep apnea hypopnea syndrome, Sleep fragmentation, Spectral content, Spontaneous arousal

Amigo, L. E., Fernandez, Q., Giralt, X., Casals, A., Amat, J., (2012). Study of patient-orthosis interaction forces in rehabilitation therapies IEEE Conference Publications 4th IEEE RAS & EMBS International Conference on Biomedical Robotics and Biomechatronics (BioRob) , IEEE (Roma, Italy) , 1098-1103

The design of mechanical joints that kinematically behave as their biological counterparts is a challenge that if not addressed properly can cause inadequate forces transmission between robot and patient. This paper studies the interaction forces in rehabilitation therapies of the elbow joint. To measure the effect of orthosis-patient misalignments, a force sensor with a novel distributed architecture has been designed and used for this study. A test-bed based on an industrial robot acting as a virtual exoskeleton that emulates the action of a therapist has been developed and the interaction forces analyzed.

Keywords: Force, Force measurement, Force sensors, Joints, Medical treatment, Robot sensing systems, Force sensors, Medical robotics, Patient rehabilitation, Biological counterparts, Distributed architecture, Elbow joint, Force sensor, Inadequate forces transmission, Industrial robot, Mechanical joints design, Orthosis-patient misalignments, Patient-orthosis interaction forces, Rehabilitation therapies, Robot, Test-bed, Virtual exoskeleton

Simao, C., Mas-Torrent, M., Crivillers, N., Lloveras, V., Artés, Juan Manuel, Gorostiza, Pau, Veciana, Jaume, Rovira, C., (2011). A robust molecular platform for non-volatile memory devices with optical and magnetic responses Nature Chemistry , 3, (5), 359-364

Bistable molecules that behave as switches in solution have long been known. Systems that can be reversibly converted between two stable states that differ in their physical properties are particularly attractive in the development of memory devices when immobilized in substrates. Here, we report a highly robust surface-confined switch based on an electroactive, persistent organic radical immobilized on indium tin oxide substrates that can be electrochemically and reversibly converted to the anion form. This molecular bistable system behaves as an extremely robust redox switch in which an electrical input is transduced into optical as well as magnetic outputs under ambient conditions. The fact that this molecular surface switch, operating at very low voltages, can be patterned and addressed locally, and also has exceptionally high long-term stability and excellent reversibility and reproducibility, makes it a very promising platform for non-volatile memory devices.

Keywords: Self-assembled monolayers, Chromophore-based monolayers, Ultrathin platinum films, Carbon free-radicals, Per-million levels, Polychlorotriphenylmethyl radicals, Electron-transfer, Surface, Logic, Quantification

Trepat, X., Fredberg, J. J., (2011). Plithotaxis and emergent dynamics in collective cellular migration Trends in Cell Biology 21, (11), 638-646

For a monolayer sheet to migrate cohesively, it has long been suspected that each constituent cell must exert physical forces not only upon its extracellular matrix but also upon neighboring cells. The first comprehensive maps of these distinct force components reveal an unexpected physical picture. Rather than showing smooth and systematic variation within the monolayer, the distribution of physical forces is dominated by heterogeneity, both in space and in time, which emerges spontaneously, propagates over great distances, and cooperates over the span of many cell bodies. To explain the severe ruggedness of this force landscape and its role in collective cell guidance, the well known mechanisms of chemotaxis, durotaxis, haptotaxis are clearly insufficient. In a broad range of epithelial and endothelial cell sheets, collective cell migration is governed instead by a newly discovered emergent mechanism of innately collective cell guidance - plithotaxis.

Keywords: Positional information, Drosophila embryo, Sheet migration, Dpp gradient, Cells, Force, Morphogenesis, Transition, Identification, Proliferation

Gauthier, Nils C., Fardin, Marc Antoine, Roca-Cusachs, Pere, Sheetz, Michael P., (2011). Temporary increase in plasma membrane tension coordinates the activation of exocytosis and contraction during cell spreading Proceedings of the National Academy of Sciences of the United States of America 108, (35), 14467-14472

Cell migration and spreading involve the coordination of membrane trafficking, actomyosin contraction, and modifications to plasma membrane tension and area. The biochemical or biophysical basis for this coordination is however unknown. In this study, we show that during cell spreading, lamellipodia protrusion flattens plasma membrane folds and blebs and, once the plasma membrane area is depleted, there is a temporary increase in membrane tension by over twofold that is followed by activation of exocytosis and myosin contraction. Further, an artificial increase in plasma membrane tension stopped lamellipodia protrusion and activated an exocytotic burst. Subsequent decrease in tension restored spreading with activation of contraction. Conversely, blebbistatin inhibition of actomyosin contraction resulted in an even greater increase in plasma membrane tension and exocytosis activation. This spatio-temporal synchronization indicates that membrane tension is the signal that coordinates membrane trafficking, actomyosin contraction, and plasma membrane area change. We suggest that cells use plasma membrane tension as a global physical parameter to control cell motility.

Keywords: Surface-area regulation, Cytoskeleton adhesion, Erythrocyte-membrane, Extensional flow, Elastic tether, Force

Melchels, Ferry P. W., Tonnarelli, Beatrice, Olivares, Andy L., Martin, Ivan, Lacroix, Damien, Feijen, Jan, Wendt, David J., Grijpma, Dirk W., (2011). The influence of the scaffold design on the distribution of adhering cells after perfusion cell seeding Biomaterials 32, (11), 2878-2884

In natural tissues, the extracellular matrix composition, cell density and physiological properties are often non-homogeneous. Here we describe a model system, in which the distribution of cells throughout tissue engineering scaffolds after perfusion seeding can be influenced by the pore architecture of the scaffold. Two scaffold types, both with gyroid pore architectures, were designed and built by stereolithography: one with isotropic pore size (412 ± 13 [mu]m) and porosity (62 ± 1%), and another with a gradient in pore size (250-500 [mu]m) and porosity (35%-85%). Computational fluid flow modelling showed a uniform distribution of flow velocities and wall shear rates (15-24 s-1) for the isotropic architecture, and a gradient in the distribution of flow velocities and wall shear rates (12-38 s-1) for the other architecture. The distribution of cells throughout perfusion-seeded scaffolds was visualised by confocal microscopy. The highest densities of cells correlated with regions of the scaffolds where the pores were larger, and the fluid velocities and wall shear rates were the highest. Under the applied perfusion conditions, cell deposition is mainly determined by local wall shear stress, which, in turn, is strongly influenced by the architecture of the pore network of the scaffold.

Keywords: Scaffolds, Microstructure, Cell adhesion, Confocal microscopy, Image analysis, Computational fluid dynamics

Sjoberg, B. M., Torrents, E., (2011). Shift in ribonucleotide reductase gene expression in pseudomonas aeruginosa during infection Infection and Immunity , 79, (7), 2663-2669

The roles of different ribonucleotide reductases (RNRs) in bacterial pathogenesis have not been studied systematically. In this work we analyzed the importance of the different Pseudomonas aeruginosa RNRs in pathogenesis using the Drosophila melanogaster host-pathogen interaction model. P. aeruginosa codes for three different RNRs with different environmental requirements. Class II and III RNR chromosomal mutants exhibited reduced virulence in this model. Translational reporter fusions of RNR gene nrdA, nrdJ, or nrdD to the green fluorescent protein were constructed to measure the expression of each class during the infection process. Analysis of the P. aeruginosa infection by flow cytometry revealed increased expression of nrdJ and nrdD and decreased nrdA expression during the infection process. Expression of each RNR class fits with the pathogenicities of the chromosomal deletion mutants. An extended understanding of the pathogenicity and physiology of P. aeruginosa will be important for the development of novel drugs against infections in cystic fibrosis patients.

Keywords: Broad-host-range, Anaerobic growth, Drosophila-melanogaster, Bacterial biofilms, Escherichia-coli, Cystic-fibrosis, Model host, Virulence, Promoter, Vectors

Pedro, L., Banos, R. C., Aznar, S., Madrid, C., Balsalobre, C., Juarez, A., (2011). Antibiotics shaping bacterial genome: Deletion of an IS91 flanked virulence determinant upon exposure to subinhibitory antibiotic concentrations PLoS ONE 6, (11), 11

The nucleoid-associated proteins Hha and YdgT repress the expression of the toxin a-hemolysin. An Escherichia coli mutant lacking these proteins overexpresses the toxin a-hemolysin encoded in the multicopy recombinant plasmid pANN202-312R. Unexpectedly, we could observe that this mutant generated clones that no further produced hemolysin (Hly(-)). Generation of Hly(-) clones was dependent upon the presence in the culture medium of the antibiotic kanamycin (km), a marker of the hha allele (hha::Tn5). Detailed analysis of different Hly(-) clones evidenced that recombination between partial IS91 sequences that flank the hly operon had occurred. A fluctuation test evidenced that the presence of km in the culture medium was underlying the generation of these clones. A decrease of the km concentration from 25 mg/l to 12.5 mg/l abolished the appearance of Hly(-) derivatives. We considered as a working hypothesis that, when producing high levels of the toxin (combination of the hha ydgT mutations with the presence of the multicopy hemolytic plasmid pANN202-312R), the concentration of km of 25 mg/l resulted subinhibitory and stimulated the recombination between adjacent IS91 flanking sequences. To further test this hypothesis, we analyzed the effect of subinhibitory km concentrations in the wild type E. coli strain MG1655 harboring the parental low copy number plasmid pHly152. At a km concentration of 5 mg/l, subinhibitory for strain MG1655 (pHly152), generation of Hly(-) clones could be readily detected. Similar results were also obtained when, instead of km, ampicillin was used. IS91 is flanking several virulence determinants in different enteric bacterial pathogenic strains from E. coli and Shigella. The results presented here evidence that stress generated by exposure to subinhibitory antibiotic concentrations may result in rearrangements of the bacterial genome. Whereas some of these rearrangements may be deleterious, others may generate genotypes with increased virulence, which may resume infection.

Keywords: Promotes horizontal dissemination, Enterica serovar typhimurium, Escherichia-coli strains, Insertion-sequence IS91, H-NS, Adaptive amplification, Pathogenicity islands, Hemolysin

Miranda Coelho, Nuno, Gonzalez-Garcia, Cristina, Salmeron-Sanchez, Manuel, Altankov, George, (2011). Arrangement of type IV collagen and laminin on substrates with controlled density of -OH groups Tissue Engineering Part A , 17, (17-18), 2245-2257

Collagen IV (Col IV) and laminin (Lam) are the main structural components of the basement membrane where they form two overlapping polymeric networks. We studied the adsorption pattern of these proteins on five model surfaces with tailored density of -OH groups obtained by copolymerization of different ratios ethyl acrylate (EA) and hydroxyl EA (HEA): X(OH) = 0, X(OH) = 0.3, X(OH) = 0.5, X(OH) = 0.7, and X(OH) = 1 (where X refers the ratio of HEA). Atomic force microscopy revealed substratum-specific adsorption patterns of Col IV and Lam, ranging from single molecules deposition on more hydrophilic substrata to the formation of complex networks on hydrophobic ones. Human umbilical endothelial cells were used to study the biological performance of adsorbed proteins, following the overall cell morphology, the quantities for cell adhesion and spreading, and the development of focal adhesion complexes and actin cytoskeleton. Surprisingly, two optima in the cellular interaction were observed-one on the most hydrophilic X(OH) = 1 and other on the relatively hydrophobic X(OH) = 0.3 substrate-valid for both Col IV and Lam. When the proteins were adsorbed consecutively, a hydrophobic shift to X(OH) = 0 substratum was obtained. Collectively, these data suggest that varying with the density of -OH groups one can tailor the conformation and the functional activity of adsorbed basement membrane proteins.

Keywords: Atomic-force microscopy, Fibronectin adsorption, Basement-membranes, Polymer surfaces, Cell-adhesion, Biomaterials, Wettability, Fibrinogen

Krishnan, Ramaswamy, Klumpers, Darinka D., Park, Chan Y., Rajendran, Kavitha, Trepat, Xavier, van Bezu, Jan, van Hinsbergh, Victor W. M., Carman, Christopher V., Brain, Joseph D., Fredberg, Jeffrey J., Butler, James P., van Nieuw Amerongen, Geerten P., (2011). Substrate stiffening promotes endothelial monolayer disruption through enhanced physical forces American Journal of Physiology - Cell Physiology , 300, (1), C146-C154

A hallmark of many, sometimes life-threatening, inflammatory diseases and disorders is vascular leakage. The extent and severity of vascular leakage is broadly mediated by the integrity of the endothelial cell (EC) monolayer, which is in turn governed by three major interactions: cell-cell and cell-substrate contacts, soluble mediators, and biomechanical forces. A potentially critical but essentially uninvestigated component mediating these interactions is the stiffness of the substrate to which the endothelial monolayer is adherent. Accordingly, we investigated the extent to which substrate stiffening influences endothelial monolayer disruption and the role of cell-cell and cell-substrate contacts, soluble mediators, and physical forces in that process. Traction force microscopy showed that forces between cell and cell and between cell and substrate were greater on stiffer substrates. On stiffer substrates, these forces were substantially enhanced by a hyperpermeability stimulus (thrombin, 1 U/ml), and gaps formed between cells. On softer substrates, by contrast, these forces were increased far less by thrombin, and gaps did not form between cells. This stiffness-dependent force enhancement was associated with increased Rho kinase activity, whereas inhibition of Rho kinase attenuated baseline forces and lessened thrombin-induced inter-EC gap formation. Our findings demonstrate a central role of physical forces in EC gap formation and highlight a novel physiological mechanism. Integrity of the endothelial monolayer is governed by its physical microenvironment, which in normal circumstances is compliant but during pathology becomes stiffer.

Keywords: Contraction, Human umbilical vein endothelial cells, Permeability, Traction force, Cell-cell contact, Cell-substrate contact, Substrate stiffness, Rho kinase, Vascular endothelial cadherin, Thrombin

Lacroix, Damien, Ramirez Patino, Juan Fernando, (2011). Finite Element Analysis of Donning Procedure of a Prosthetic Transfemoral Socket Annals of Biomedical Engineering , 39, (12), 2972-2983

Lower limb amputation is a severe psychological and physical event in a patient. A prosthetic solution can be provided but should respond to a patient-specific need to accommodate for the geometrical and biomechanical specificities. A new approach to calculate the stress-strain state at the interaction between the socket and the stump of five transfemoral amputees is presented. In this study the socket donning procedure is modeled using an explicit finite element method based on the patient-specific geometry obtained from CT and laser scan data. Over stumps the mean maximum pressure is 4 kPa (SD 1.7) and the mean maximum shear stresses are 1.4 kPa (SD 0.6) and 0.6 kPa (SD 0.3) in longitudinal and circumferential directions, respectively. Locations of the maximum values are according to pressure zones at the sockets. The stress-strain states obtained in this study can be considered more reliable than others, since there are normal and tangential stresses associated to the socket donning procedure.

Keywords: Trans-tibial prosthesis, Knee residual limb, Pressure distribution, Transtibial amputees, Stump/socket interface, Mechanical conditions, Load-transfer, Soft-tissues, Stresses, Contact

Hristova, K., Pecheva, E., Pramatarova, L., Altankov, G., (2011). Improved interaction of osteoblast-like cells with apatite-nanodiamond coatings depends on fibronectin Journal of Materials Science: Materials in Medicine , 22, (8), 1891-1900

New apatite (AP)/nanodiamond (ND) coating has been developed to improve physical and biological properties of stainless steel (SS) versus single AP coating. Homogeneously electrodeposited AP-ND layer demonstrates increased mechanical strength, interlayer cohesion and ductility. In the absence of serum, osteoblast-like MG63 cells attach well but poorly spread on both AP and AP-ND substrata. Pre-adsorption with serum or fibronectin (FN) improves the cellular interaction-an effect that is better pronounced on the AP-ND coating. In single protein adsorption study fluorescein isothiocyanate-labeled FN (FITC-FN) shows enhanced deposition on the AP-ND layer consistent with the significantly improved cell adhesion, spreading and focal adhesions formation (in comparison to SS and AP), particularly at low FN adsorption concentrations (1 mu g/ml). Higher FN concentrations (20 mu g/ml) abolish this difference suggesting that the promoted cellular interaction of serum (where FN is low) is caused by the greater affinity for FN. Moreover, it is found that MG63 cells tend to rearrange both adsorbed and secreted FN on the AP-ND layer suggesting facilitated FN matrix formation.

Keywords: Extracellular-matrix, Protein adsorption, Integrins, Adhesion, Biomaterials, Surfaces, Polymerization, Composite, Implants, Titanium

Moore, S. W., Roca-Cusachs, P., Sheetz, M. P., (2010). Stretchy proteins on stretchy substrates: The important elements of integrin-mediated rigidity sensing Developmental Cell 19, (2), 194-206

Matrix and tissue rigidity guides many cellular processes, including the differentiation of stem cells and the migration of cells in health and disease. Cells actively and transiently test rigidity using mechanisms limited by inherent physical parameters that include the strength of extracellular attachments, the pulling capacity on these attachments, and the sensitivity of the mechanotransduction system. Here, we focus on rigidity sensing mediated through the integrin family of extracellular matrix receptors and linked proteins and discuss the evidence supporting these proteins as mechanosensors.

Keywords: Focal adhesion kinase, Atomic Force Microscopy, Smooth-muscle cells, Traction forces, Living cells, Mechanical force, Locomoting cells

Coelho, N. M., Gonzalez-Garcia, C., Planell, J. A., Salmeron-Sanchez, M., Altankov, G., (2010). Different assembly of type iv collagen on hydrophilic and hydrophobic substrata alters endothelial cells interaction European Cells & Materials , 19, 262-272

Considering the structural role of type IV collagen (Col IV) in the assembly of the basement membrane (BM) and the perspective of mimicking its organization for vascular tissue engineering purposes, we studied the adsorption pattern of this protein on model hydrophilic (clean glass) and hydrophobic trichloro(octadecyl) silane (ODS) surfaces known to strongly affect the behavior of other matrix proteins. The amount of fluorescently labeled Col IV was quantified showing saturation of the surface for concentration of the adsorbing solution of about 50 mu g/ml, but with approximately twice more adsorbed protein on ODS. AFM studies revealed a fine-nearly single molecular size-network arrangement of Col IV on hydrophilic glass, which turns into a prominent and growing polygonal network consisting of molecular aggregates on hydrophobic ODS. The protein layer forms within minutes in a concentration-dependent manner. We further found that human umbilical vein endothelial cells (HUVEC) attach less efficiently to the aggregated Col IV (on ODS), as judged by the significantly altered cell spreading, focal adhesions formation and the development of actin cytoskeleton. Conversely, the immunofluorescence studies for integrins revealed that the fine Col IV network formed on hydrophilic substrata is better recognized by the cells via both alpha 1 and alpha 2 heterodimers which support cellular interaction, apart from these on hydrophobic ODS where almost no clustering of integrins was observed.

Keywords: Collagen type IV, Adsorption, Assembly, Hydrophilic, Hydrophobic, Surfaces

Valente, T., Gella, A., Fernàndez-Busquets, X., Unzeta, M., Durany, N., (2010). Immunohistochemical analysis of human brain suggests pathological synergism of Alzheimer's disease and diabetes mellitus Neurobiology of Disease , 37, (1), 67-76

It has been extensively reported that diabetes mellitus (DM) patients have a higher risk of developing Alzheimer's disease (AD). but a mechanistic connection between both pathologies has not been provided so far Carbohydrate-derived advanced glycation endproducts (AGEs) have been implicated in the chronic complications of DM and have been reported to play an important role in the pathogenesis of AD. The earliest histopathological manifestation of AD is the apparition of extracellular aggregates of the amyloid beta peptide (A beta). To investigate possible correlations between AGEs and A beta aggregates with both pathologies. we have performed an immuhistochemical study in human post-mortem samples of AD, AD with diabetes (ADD). diabetic and nondemented controls ADD brains showed increased number of A beta dense plaques and receptor for AGEs (RACE)-positive and Tau-positive cells, higher AGEs levels and major microglial activation, compared to AD brain. Our results indicate that ADD patients present a significant increase of cell damage through a RAGE-dependent mechanism, suggesting that AGEs may promote the generation of an oxidative stress vicious cycle, which can explain the severe progression of patients with both pathologies.

Keywords: Abeta, Alzheimer's disease, Rage, Ages, Diabetes, Immunohistochemistry, Advanced glycation endproducts, Beta-amyloid peptide, End-products, Oxidative stress, Advanced glycosylation, Synaptic dysfunction, Cross-linking

Fernàndez-Busquets, X., Ponce, J., Bravo, R., Arimon, M., Martianez, T., Gella, A., Cladera, J., Durany, N., (2010). Modulation of amyloid beta peptide(1-42) cytotoxicity and aggregation in vitro by glucose and chondroitin sulfate Current Alzheimer Research , 7, (5), 428-438

One mechanism leading to neurodegeneration during Alzheimer's Disease (AD) is amyloid beta peptide (A beta)-induced neurotoxicity. Among the factors proposed to potentiate A beta toxicity is its covalent modification through carbohydrate-derived advanced glycation endproducts (AGEs). Other experimental evidence, though, indicates that certain polymeric carbohydrates like the glycosaminoglycan (GAG) chains found in proteoglycan molecules attenuate the neurotoxic effect of A beta in primary neuronal cultures. Pretreatment of the 42-residue A beta fragment (A beta(1-42)) with the ubiquitous brain carbohydrates, glucose, fructose, and the GAG chondroitin sulfate B (CSB) inhibits A beta beta(1-42)-induced apoptosis and reduces the peptide neurotoxicity on neuroblastoma cells, a cytoprotective effect that is partially reverted by AGE inhibitors such as pyridoxamine and L-carnosine. Thioflavin T fluorescence measurements indicate that at concentrations close to physiological, only CSB promotes the formation of A beta amyloid fibril structure. Atomic force microscopy imaging and Western blot analysis suggest that glucose favours the formation of globular oligomeric structures derived from aggregated species. Our data suggest that at short times carbohydrates reduce A beta(1-42) toxicity through different mechanisms both dependent and independent of AGE formation.

Keywords: Alzheimer's disease, Advanced glycation endproducts, Amyloid fibrils, Amyloid beta peptide, Apoptosis, Carbohydrates, Glycosaminoglycans

Comelles, J., Estevez, M., Martinez, E., Samitier, J., (2010). The role of surface energy of technical polymers in serum protein adsorption and MG-63 cells adhesion Nanomedicine: Nanotechnology Biology and Medicine , 6, (1), 44-51

Polymeric materials are widely used as supports for cell culturing in medical implants and as scaffolds for tissue regeneration. However, novel applications in the biosensor field require materials to be compatible with cell growth and at the same time be suitable for technological processing. Technological polymers are key materials in the fabrication of disposable parts and other sensing elements. As such, it is essential to characterize the surface properties of technological polymers, especially after processing and sterilization. It is also important to understand how technological polymers affect cell behavior when in contact with polymer materials. Therefore, the aim of this research was to study how surface energy and surface roughness affect the biocompatibility of three polymeric materials widely used in research and industry: poly (methyl methacrylate), polystyrene, and poly(dimethylsiloxane). Glass was used as the control material. From the Clinical Editor: Polymeric materials are widely used as supports for cell culturing in medical implants and as scaffolds for tissue regeneration. The aim of this research is to study how surface energy and surface roughness affect the biocompatibility of three polymeric materials widely used in research and industry: poly(methylmethacrylate) (PMMA), polystyrene (PS), and poly(dimethylsiloxane) (PDMS).

Keywords: Thin-films, Poly(methyl methacrylate), Osteoblast adhesion, Electron-microscopy, Fibronectin, Polystyrene, Oly(dimethylsiloxane), Biocompatibility, Hydroxyapatite, Behavior

Estevez, M., Fernandez-Ulibarri, I., Martinez, E., Egea, G., Samitier, J., (2010). Changes in the internal organization of the cell by microstructured substrates Soft Matter 6, (3), 582-590

Surface features at the micro and nanometre scale have been shown to influence and even determine cell behaviour and cytoskeleton organization through direct mechanotransductive pathways. Much less is known about the function and internal distribution of organelles of cells grown on topographically modified surfaces. In this study, the nanoimprint lithography technique was used to manufacture poly(methyl methacrylate) (PMMA) sheets with a variety of features in the micrometre size range. Normal rat kidney (NRK) fibroblasts were cultured on these substrates and immunofluorescence staining assays were performed to visualize cell adhesion, the organization of the cytoskeleton and the morphology and subcellular positioning of the Golgi complex. The results show that different topographic features at the micrometric scale induce different rearrangements of the cell cytoskeleton, which in turn alter the positioning and morphology of the Golgi complex. Microposts and microholes alter the mechanical stability of the Golgi complex by modifying the actin cytoskeleton organization leading to the compaction of the organelle. These findings prove that physically modified surfaces are a valuable tool with which to study the dynamics of cell cytoskeleton organization and its subsequent repercussion on internal cell organization and associated function.

Keywords: Actin stress fibers, Golgi-complex, Focal adhesions, Cytoskeletal organization, Osteoblast adhesion, Mammalian-cells, Micron-scale, Nanoscale, Dynamics, Rho

Madronal, Noelia, Lopez-Aracil, Cristina, Rangel, Alejandra, del Rio, Jose A., Delgado-Garcia, Jose M., Gruart, Agnes, (2010). Effects of Enriched Physical and Social Environments on Motor Performance, Associative Learning, and Hippocampal Neurogenesis in Mice PLoS ONE 5, (6), e11130

We have studied the motor abilities and associative learning capabilities of adult mice placed in different enriched environments. Three-month-old animals were maintained for a month alone (AL), alone in a physically enriched environment (PHY), and, finally, in groups in the absence (SO) or presence (SOPHY) of an enriched environment. The animals' capabilities were subsequently checked in the rotarod test, and for classical and instrumental learning. The PHY and SOPHY groups presented better performances in the rotarod test and in the acquisition of the instrumental learning task. In contrast, no significant differences between groups were observed for classical eyeblink conditioning. The four groups presented similar increases in the strength of field EPSPs (fEPSPs) evoked at the hippocampal CA3-CA1 synapse across classical conditioning sessions, with no significant differences between groups. These trained animals were pulse-injected with bromodeoxyuridine (BrdU) to determine hippocampal neurogenesis. No significant differences were found in the number of NeuN/BrdU double-labeled neurons. We repeated the same BrdU study in one-month-old mice raised for an additional month in the above-mentioned four different environments. These animals were not submitted to rotarod or conditioned tests. Non-trained PHY and SOPHY groups presented more neurogenesis than the other two groups. Thus, neurogenesis seems to be related to physical enrichment at early ages, but not to learning acquisition in adult mice.

Keywords: Long-term potentiation, Adult neurogenesis, Synaptic transmission, Cell proliferation, CA3-CA1 synapse, Granule cells

Lagunas, A., Comelles, J., Martinez, E., Samitier, J., (2010). Universal chemical gradient platforms using poly(methyl methacrylate) based on the biotin streptavidin interaction for biological applications Langmuir 26, (17), 14154-14161

This article describes a simple method for the construction of a universal surface chemical gradient platform based on the biotin streptavidin model. In this approach, surface chemical gradients were prepared in poly(methyl methacrylate) (PM MA), a biocompatible polymer, by a controlled hydrolysis procedure. The physicochemical properties of the resulting modified surfaces were extensively characterized. Chemical analysis carried out via time-of-flight secondary ion mass spectrometry (ToRSIMS) and X-ray photoelectron spectroscopy (XPS) showed the formation of a smooth, highly controllable carboxylic acid gradient of increasing concentration along the sample surface. Atomic force microscopy (AFM) and contact angle (CA) results indicate that, in contrast with most of the chemical gradient methods published in the literature, the chemical modification of the polymer surface barely affects its physical properties. The introduction of carboxylic acid functionality along the surface was then used for biomolecule anchoring. For this purpose, the surface was activated and derivatized first with biotin and finally with streptavidin (SA V) in a directed orientation fashion. The SAV gradient was qualitatively assessed by fluorescence microscopy analysis and quantified by surface plasmon resonance (SPR) in order to establish a quantitative relationship between SAV surface densities and the surface location. The usefulness of the fabrication method described for biological applications was tested by immobilizing biotinylated bradykinin onto the SAV gradient. This proof-of-concept application shows the effectiveness of the concentration range of the gradient because the effects of bradykinin on cell morphology were observed to increase gradually with increasing drug concentrations. The intrinsic characteristics of the fabricated gradient platform (absence of physicochemical modifications other than those due to the biomolecules included) allow us to attribute cell behavior unequivocally to the biomolecule surface density changes.

Keywords: Wettability gradient, Polyethylene surface, Combinatorial, Immobilization, Biomaterials, Fabrication, Deposition, Bradykinin, Monolayers, Discharge

Toromanov, Georgi, González-García, Cristina, Altankov, George, Salmerón-Sánchez, Manuel, (2010). Vitronectin activity on polymer substrates with controlled -OH density Polymer , 51, (11), 2329-2336

Vitronectin (VN) adsorption on a family of model substrates consisting of copolymers of ethyl acrylate and hydroxyl ethylacrylate in different ratios (to obtain a controlled surface density of -OH groups) was investigated by Atomic Force Microscopy (AFM). It is shown that the fraction of the substrate covered by the protein depends strongly on the amount of hydroxyl groups in the sample and it monotonically decreases as the -OH density increases. Isolated globular-like VN molecules are observed on the surfaces with the higher OH density. As the fraction of hydroxyl groups decreases, aggregates of 3-5 VN molecules are observed on the sample. Overall cell morphology, focal adhesion formation and actin cytoskeleton development are investigated to assess the biological activity of the adsorbed VN on the different surfaces. Dermal fibroblast cells show excellent material interaction on the more hydrophobic samples (OH contents lower than 0.5), which reveals enhanced VN activity on this family of substrates as compared with other extracellular matrix proteins (e.g., fibronectin and fibrinogen).

Keywords: Copolymers, Vitronectin, AFM, Self-assembled monolayers, Cell-adhesion, Thermal transitions, Protein adsorption, Surfaces, Fibronectin, Biomaterials, Attachment, Fibrinogen

Gugutkov, D., Altankov, G., Hernandez, J. C. R., Pradas, M. M., Sanchez, M. S., (2010). Fibronectin activity on substrates with controlled -OH density Journal of Biomedical Materials Research - Part A , 92A, (1), 322-331

Adhesion of human fibroblast to a family of fibronectin (FN) coated model substrates consisting of copolymers of ethyl acrylate and hydroxyl ethylacrylate in different ratios to obtain a controlled surface density of -OH groups was investigated. Cell adhesion and spreading surprisingly decreased as the fraction of -OH groups on the Surface increased. AFM studies of FN conformation revealed formation of a protein network on the more hydrophobic surfaces. The density of this network diminished as the fraction of -OH groups in the sample increased, up to a maximal -OH concentration at which, instead of the network, only IN aggregates were observed. The kinetics of network development was followed at different adsorption times. Immunofluorescence for vinculin revealed the formation of well-developed focal adhesion complexes on the more hydrophobic surface (similar to the control glass), which became less defined as the fraction of -OH groups increased. Thus, the efficiency of cell adhesion is enhanced by the formation of FN networks on the substrate, directly revealing the importance of the adsorbed protein conformation for cell adhesion. However, cell-dependent reorganization of substrate-associated FN, which usually takes place on more hydrophilic substrates (as do at the control glass slides), was not observed in this system, suggesting the increased strength of protein-to-substrate interaction. Instead, the late FN matrix formation-after 3 days of culture-was again better pronounced on the more hydrophobic substrates and decreased as the fraction of -OH groups increase, which is in a good agreement with the results for overall cell morphology and focal adhesion formation.

Keywords: Cell adhesion, Fibronectin, Fibroblast, Extracellular matrix, AFM

Correa, R., Laciar, E., Arini, P., Jané, R., (2010). Analysis of QRS loop in the Vectorcardiogram of patients with Chagas' disease Engineering in Medicine and Biology Society (EMBC) 32nd Annual International Conference of the IEEE , IEEE (Buenos Aires, Argentina) , 2561-2564

In the present work, we have studied the QRS loop in the Vectorcardiogram (VCG) of 95 chronic chagasic patients classified in different groups (I, II and III) according to their degree of myocardial damage. For comparison, the VCGs of 11 healthy subjects used as control group (Group O) were also examined. The QRS loop was obtained for each patient from the XYZ orthogonal leads of their High-Resolution Electrocardiogram (HRECG) records. In order to analyze the variations of QRS loop in each detected beat, it has been proposed in this study the following vectorcardiographic parameters a) Maximum magnitude of the cardiac depolarization vector, b) Volume, c) Area of QRS loop, d) Ratio between the Area and Perimeter, e) Ratio between the major and minor axes of the QRS loop and f) QRS loop Energy. It has been found that one or more indexes exhibited statistical differences (p<0.05) between groups 0-II, O-III, I-II, I-III and II-III. We concluded that the proposed method could be use as complementary diagnosis technique to evaluate the degree of myocardial damage in chronic chagasic patients.

Keywords: Practical, Experimental/ bioelectric phenomena, Diseases, Electrocardiography, Medical signal, Processing/ QRS loop, Vectorcardiogram, Cardiac depolarization vector, Myocardial damage, Chagas disease, Complementary diagnosis technique, High-resolution electrocardiogram

Tarzan-Lorente, M., Gutierrez-Galvez, A., Martinez, D., Marco, S., (2010). A biologically inspired associative memory for artificial olfaction Practica 2010 International Joint Conference on Neural Networks (IJCNN 2010) , IEEE, Piscataway, NJ, USA (Barcelona, Spain) , 6 pp.

In this paper, we propose a biologically inspired architecture for a Hopfield-like associative memory applied to artificial olfaction. The proposed algorithm captures the projection between two neural layers of the insect olfactory system (Antennal Lobe and Mushroom Body) with a kernel based projection. We have tested its classification performance as a function of the size of the training set and the time elapsed since training and compared it with that obtained with a Support Vector Machine.

Keywords: Biocomputing, Chemioception, Content-addressable storage, Hopfield neural nets, Support vector machines

Garde, A., Sörnmo, L., Jané, R., Giraldo, B. F., (2010). Correntropy-based nonlinearity test applied to patients with chronic heart failure Engineering in Medicine and Biology Society (EMBC) 32nd Annual International Conference of the IEEE , IEEE (Buenos Aires, Argentina) , 2399-2402

In this study we propose the correntropy function as a discriminative measure for detecting nonlinearities in the respiratory pattern of chronic heart failure (CHF) patients with periodic or nonperiodic breathing pattern (PB or nPB, respectively). The complexity seems to be reduced in CHF patients with higher risk level. Correntropy reflects information on both, statistical distribution and temporal structure of the underlying dataset. It is a suitable measure due to its capability to preserve nonlinear information. The null hypothesis considered is that the analyzed data is generated by a Gaussian linear stochastic process. Correntropy is used in a statistical test to reject the null hypothesis through surrogate data methods. Various parameters, derived from the correntropy and correntropy spectral density (CSD) to characterize the respiratory pattern, presented no significant differences when extracted from the iteratively refined amplitude adjusted Fourier transform (IAAFT) surrogate data. The ratio between the powers in the modulation and respiratory frequency bands R was significantly different in nPB patients, but not in PB patients, which reflects a higher presence of nonlinearities in nPB patients than in PB patients.

Keywords: Practical, Theoretical or Mathematical, Experimental/cardiology diseases, Fourier transforms, Medical signal processing, Pattern classification, Pneumodynamics, Spectral analysis, Statistical analysis, Stochastic processes/ correntropy based nonlinearity test, Chronic heart failure, Correntropy function, Respiratory pattern nonlinearities, CHF patients, Nonperiodic breathing pattern, Dataset statistical distribution, Dataset temporal structure, Nonlinear information, Null hypothesis, Gaussian linear stochastic process, Statistical test, Correntropy spectral density, Iteratively refined amplitude adjusted Fourier transform, Surrogate data, Periodic breathing pattern

Trepat, X., Wasserman, M. R., Angelini, T. E., Millet, E., Weitz, D. A., Butler, J. P., Fredberg, J. J., (2009). Physical forces during collective cell migration Nature Physics 5, (6), 426-430

Fundamental biological processes including morphogenesis, tissue repair and tumour metastasis require collective cell motions(1-3), and to drive these motions cells exert traction forces on their surroundings(4). Current understanding emphasizes that these traction forces arise mainly in 'leader cells' at the front edge of the advancing cell sheet(5-9). Our data are contrary to that assumption and show for the first time by direct measurement that traction forces driving collective cell migration arise predominately many cell rows behind the leading front edge and extend across enormous distances. Traction fluctuations are anomalous, moreover, exhibiting broad non-Gaussian distributions characterized by exponential tails(10-12). Taken together, these unexpected findings demonstrate that although the leader cell may have a pivotal role in local cell guidance, physical forces that it generates are but a small part of a global tug-of-war involving cells well back from the leading edge.

Keywords: Focal adhesions, Granular matter, Bead packs, Morphogenesis, Sheets, Actin, Fluctuations, Fibroblasts, Microscopy, Diversity

Fernàndez-Busquets, X., Körnig, A., Bucior, I., Burger, M. M., Anselmetti, D., (2009). Self-recognition and Ca2+-dependent carbohydrate-carbohydrate cell adhesion provide clues to the cambrian explosion Molecular Biology and Evolution , 26, (11), 2551-2561

The Cambrian explosion of life was a relatively short period approximately 540 Ma that marked a generalized acceleration in the evolution of most animal phyla, but the trigger of this key biological event remains elusive. Sponges are the oldest extant Precambrian metazoan phylum and thus a valid model to study factors that could have unleashed the rise of multicellular animals. One such factor is the advent of self-/non-self-recognition systems, which would be evolutionarily beneficial to organisms to prevent germ-cell parasitism or the introduction of deleterious mutations resulting from fusion with genetically different individuals. However, the molecules responsible for allorecognition probably evolved gradually before the Cambrian period, and some other (external) factor remains to be identified as the missing triggering event. Sponge cells associate through calcium-dependent, multivalent carbohydrate-carbohydrate interactions of the g200 glycan found on extracellular proteoglycans. Single molecule force spectroscopy analysis of g200-g200 binding indicates that calcium affects the lifetime (+Ca/-Ca: 680 s/3 s) and bond reaction length (+Ca/-Ca: 3.47 /2.27). Calculation of mean g200 dissociation times in low and high calcium within the theoretical framework of a cooperative binding model indicates the nonlinear and divergent characteristics leading to either disaggregated cells or stable multicellular assemblies, respectively. This fundamental phenomenon can explain a switch from weak to strong adhesion between primitive metazoan cells caused by the well-documented rise in ocean calcium levels at the end of Precambrian time. We propose that stronger cell adhesion allowed the integrity of genetically uniform animals composed only of "self" cells, facilitating genetic constitutions to remain within the metazoan individual and be passed down inheritance lines. The Cambrian explosion might have been triggered by the coincidence in time of primitive animals endowed with self-/non-self-recognition and of a surge in seawater calcium that increased the binding forces between their calcium-dependent cell adhesion molecules.

Keywords: Calcium, Cambrian explosion, Carbohydrates, Cell adhesion, Origin of Metazoa, Sponges

Roca-Cusachs, P., Gauthier, N. C., del Rio, A., Sheetz, M. P., (2009). Clustering of alpha(5)beta(1) integrins determines adhesion strength whereas alpha(v)beta(3) and talin enable mechanotransduction Proceedings of the National Academy of Sciences of the United States of America 106, (38), 16245-16250

A key molecular link between cells and the extracellular matrix is the binding between fibronectin and integrins alpha(5)beta(1) and alpha(v)beta(3). However, the roles of these different integrins in establishing adhesion remain unclear. We tested the adhesion strength of fibronectin-integrin-cytoskeleton linkages by applying physiological nanonewton forces to fibronectin-coated magnetic beads bound to cells. We report that the clustering of fibronectin domains within 40 nm led to integrin alpha(5)beta(1) recruitment, and increased the ability to sustain force by over six-fold. This force was supported by alpha(5)beta(1) integrin clusters. Importantly, we did not detect a role of either integrin alpha(v)beta(3) or talin 1 or 2 in maintaining adhesion strength. Instead, these molecules enabled the connection to the cytoskeleton and reinforcement in response to an applied force. Thus, high matrix forces are primarily supported by clustered alpha(5)beta(1) integrins, while less stable links to alpha(v)beta(3) integrins initiate mechanotransduction, resulting in reinforcement of integrin-cytoskeleton linkages through talin-dependent bonds.

Keywords: Cell-adhesion, Mechanical force, Vinculin-binding, Fibronectin, Activation, Dynamics, Domain, Alpha-v-beta-3, Translocation, Bonds

Nicolas, O., Gavin, R., Del Rio, J. A., (2009). New insights into cellular prion protein (PrPc) functions: The "ying and yang" of a relevant protein Brain Research Reviews , 61, (2), 170-184

The conversion of cellular prion protein (PrPc) a GPI-anchored protein, into a protease-K-resistant and infective form (generally termed PrPsc) is mainly responsible for Transmissible Spongiform Encephalopathies (TSEs), characterized by neuronal degeneration and progressive loss of basic brain functions. Although PrPc is expressed by a wide range of tissues throughout the body, the complete repertoire of its functions has not been fully deter-mined. Recent studies have confirmed its participation in basic physiological processes such as cell proliferation and the regulation of cellular homeostasis. Other studies indicate that PrPc interacts with several molecules to activate signaling cascades with a high number of cellular effects. To deter-mine PrPc functions, transgenic mouse models have been generated in the last decade. In particular, mice lacking specific domains of the PrPc protein have revealed the contribution of these domains to neurodegenerative processes. A dual role of PrPc has been shown, since most authors report protective roles for this protein while others describe pro-apoptotic functions. in this review, we summarize new findings on PrPc functions, especially those related to neural degeneration and cell signaling.

Keywords: Prion, Doppel, Shadoo, Cell death, Cell proliferation, Cell differentiation

Fernandez, Javier G., Mills, C. A., Samitier, J., (2009). Complex microstructured 3D surfaces using chitosan biopolymer Small 5, (5), 614-620

A technique for producing micrometer-scale structures over large, nonplanar chitosan surfaces is described. The technique makes use of the rheological characteristics (deformability) of the chitosan to create freestanding, three-dimensional scaffolds with controlled shapes, incorporating defined microtopography. The results of an investigation into the technical limits of molding different combinations of shapes and microtopographies are presented, highlighting the versatility of the technique when used irrespectively with inorganic or delicate organic moulds. The final, replicated scaffolds presented here are patterned with arrays of one-micrometer-tall microstructures over large areas. Structural integrity is characterized by the measurement of structural degradation. Human umbilical vein endothelial cells cultured on a tubular scaffold show that early cell growth is conditioned by the microtopography and indicate possible uses for the structures in biomedical applications. For those applications requiring improved chemical and mechanical resistance, the structures can be replicated in poly(dimethyl siloxane).

Keywords: Biocompatible Materials/ chemistry, Cell Adhesion, Cell Culture Techniques/ methods, Cell Proliferation, Cells, Cultured, Chitosan/ chemistry, Crystallization/methods, Endothelial Cells/ cytology/ physiology, Humans, Materials Testing, Nanostructures/ chemistry/ ultrastructure, Nanotechnology/methods, Particle Size, Surface Properties, Tissue Engineering/methods

Diez-Ahedo, Ruth , Normanno, Davide , Esteban, Olga,, Bakker, Gert-Jan, Figdor, Carl, Cambi, Alessandra , Garcia-Parajo, M. F., (2009). Dynamic re-organization of individual adhesion nanoclusters in living cells by ligand-patterned surfaces Small 5, (11), 1258-1263

Ligand-patterned surfaces alter the spatio-temporal organization of specific receptors on the cell membrane. Chemically confined surfaces are fabricated using microcontact patterning. The dynamic re-organization of the integrin LFA-1 in living cells is monitored at the single-molecule level using total internal reflection fluorescence. The image on the left shows individual LFA-1 nanoclusters on a single cell being recruited to ligand-rich areas of the pattern.

Keywords: Cell adhesion, Microcontact printing, Patterning, Single molecule studies

Carreras, A., Almendros, I., Acerbi, I., Montserrat, J. M., Navajas, D., Farre, R., (2009). Obstructive apneas induce early release of mesenchymal stem cells into circulating blood Sleep , 32, (1), 117-119

STUDY OBJECTIVES: To investigate whether noninvasive application of recurrent airway obstructions induces early release of mesenchymal stem cells into the circulating blood in a rat model of obstructive sleep apnea. DESIGN: Prospective controlled animal study. SETTING: University laboratory. PATIENTS OR PARTICIPANTS: Twenty male Sprague-Dawley rats (250-300 g). INTERVENTIONS: A specially designed nasal mask was applied to the anesthetized rats. Ten rats were subjected to a pattern of recurrent obstructive apneas (60 per hour, lasting 15 seconds each) for 5 hours. Ten anesthetized rats were used as controls. MEASUREMENTS AND RESULTS: Mesenchymal stem cells from the blood and bone marrow samples were isolated and cultured to count the total number of colony-forming unit fibroblasts (CFU-F) of adherent cells after 9 days in culture. The number of CFU-F from circulating blood was significantly (P = 0.02) higher in the rats subjected to recurrent obstructive apneas (5.00 +/- 1.16; mean +/- SEM) than in controls (1.70 +/- 0.72). No significant (P = 0.54) differences were observed in CFU-F from bone marrow. CONCLUSIONS: Application of a pattern of airway obstructions similar to those experienced by patients with sleep apnea induced an early mobilization of mesenchymal stem cells into circulating blood.

Keywords: Adipocytes/cytology, Animals, Blood Cell Count, Bone Marrow Cells/ cytology, Cell Adhesion/physiology, Cell Count, Cell Differentiation/physiology, Cell Division/physiology, Disease Models, Animal, Fibroblasts/cytology, Male, Mesenchymal Stem Cells/ cytology, Osteocytes/cytology, Rats, Rats, Sprague-Dawley, Sleep Apnea, Obstructive/ blood, Stem Cells/cytology

Gugutkov, Dencho, Gonzalez-Garcia, Cristina, Rodriguez Hernandez, Jose Carlos, Altankov, George, Salmeron-Sanchez, Manuel, (2009). Biological activity of the substrate-induced fibronectin network: insight into the third dimension through electrospun fibers Langmuir 25, (18), 10893-10900

Fibronectin (FN) fibrillogenesis is a cell-mediated process involving integrin activation that results in conformational changes of FN molecules and the organization of actin cytoskeleton. A similar process can be induced by some chemistries in the absence of cells, e.g., poly(ethyl acrylate) (PEA), which enhance FN-FN interactions leading to the formation of a biologically active network. Atomic force microscopy images of single FN molecules, at the early stages of adsorption on plane PEA, allow one to rationalize the process. Further, the role of the spatial organization of the FN network on the cellular response is investigated through its adsorption on electrospun fibers. Randomly oriented and aligned PEA fibers were prepared to mimic the three-dimensional organization of the extracellular matrix. The formation of the FN network on the PEA fibers but not on the supporting coverglass was confirmed. Fibroblasts aligned with oriented fibers, displayed extended morphology, developed linearly organized focal adhesion complexes, and matured actin filaments. Conversely, on random PEA fibers, cells acquired polygonal morphology with altered actin cytoskeleton but well-developed focal adhesions. Late FN matrix formation was also influenced: spatially organized FN matrix fibrils along the oriented PEA fibers and an altered arrangement on random ones.

Keywords: AFM, Cell-adhesion, Dependent conformations, Hydrophobic surfaces, Extracellular-matrix, Bound fibronectin, Polymer surfaces, Integrin binding, Biocompatibility, Adsorption

Gimenez-Oya, V., Villacanas, O., Fernàndez-Busquets, X., Rubio-Martinez, J., Imperial, S., (2009). Mimicking direct protein-protein and solvent-mediated interactions in the CDP-methylerythritol kinase homodimer: a pharmacophore-directed virtual screening approach Journal of Molecular Modeling , 15, (8), 997-1007

The 2C-methylerythritol 4-phosphate (MEP) pathway for the biosynthesis of isopentenyl pyrophosphate and its isomer dimethylallyl pyrophosphate, which are the precursors of isoprenoids, is present in plants, in the malaria parasite Plasmodium falciparum and in most eubacteria, including pathogenic agents. However, the MEP pathway is absent from fungi and animals, which have exclusively the mevalonic acid pathway. Given the characteristics of the MEP pathway, its enzymes represent potential targets for the generation of selective antibacterial, antimalarial and herbicidal molecules. We have focussed on the enzyme 4-(cytidine 5'-diphospho)-2-C-methyl-D: -erythritol kinase (CMK), which catalyses the fourth reaction step of the MEP pathway. A molecular dynamics simulation was carried out on the CMK dimer complex, and protein-protein interactions analysed, considering also water-mediated interactions between monomers. In order to find small molecules that bind to CMK and disrupt dimer formation, interactions observed in the dynamics trajectory were used to model a pharmacophore used in database searches. Using an intensity-fading matrix-assisted laser desorption/ionisation time-of-flight mass spectrometry approach, one compound was found to interact with CMK. The data presented here indicate that a virtual screening approach can be used to identify candidate molecules that disrupt the CMK-CMK complex. This strategy can contribute to speeding up the discovery of new antimalarial, antibacterial, and herbicidal compounds.

Keywords: Solvent-mediated interactions, Protein-protein interactions, Molecular dynamics, Drug design, Intensisty-fading MALDI-TOF mass spectrometry

Rico, P., Rodriguez Hernandez, J. C., Moratal, D., Altankov, G., Monleon Pradas, M., Salmeron-Sanchez, M., (2009). Substrate-induced assembly of fibronectin into networks. Influence of surface chemistry and effect on osteoblast adhesion Tissue Engineering Part A , 15, (00), 1-11

The influence of surface chemistry -substrates with controlled surface density of -OH groups- on fibronectin conformation and distribution is directly observed by Atomic Force Microscopy (AFM). FN fibrillogenesis, which is known to be a process triggered by interaction with integrins, is shown in our case to be induced by the substrate (in absence of cells), which is able to enhance FN-FN interactions leading to the formation of a protein network on the material surface. This phenomenon depends both on surface chemistry and protein concentration. The level of the FN fibrillogenesis was quantified by calculating the fractal dimension of the adsorbed protein from image analysis of the AFM results. The total amount of adsorbed FN is obtained by making use of a methodology which employs western-blotting combined with image analysis of the corresponding protein bands, with the lowest sensitivity threshold equal to 15 ng of adsorbed protein. Furthermore, FN adsorption is correlated to human osteoblast adhesion through morphology and actin cytoskeleton formation. Actin polymerization is in need of the formation of the protein network on the substrate's surface. Cell morphology is more rounded (as quantified by calculating the circularity of the cells by image analysis) the lower the degree of FN fibrillogenesis on the substrate.

Keywords: Cell-adhesion, Conformational-changes, Electron-microscopy, Protein adsorption, Fractal dimension, Integrin binding, Biocompatibility, Monolayers, Matrix, Fibrillogenesis

Kirchhof, K., Hristova, K., Krasteva, N., Altankov, G., Groth, T., (2009). Multilayer coatings on biomaterials for control of MG-63 osteoblast adhesion and growth Journal of Materials Science: Materials in Medicine , 20, (4), 897-907

Here, the layer-by-layer technique (LbL) was used to modify glass as model biomaterial with multilayers of chitosan and heparin to control the interaction with MG-63 osteoblast-like cells. Different pH values during multilayer formation were applied to control their physico-chemical properties. In the absence of adhesive proteins like plasma fibronectin (pFN) both plain layers were rather cytophobic. Hence, the preadsorption of pFN was used to enhance cell adhesion which was strongly dependent on pH. Comparing the adhesion promoting effects of pFN with an engineered repeat of the FN III fragment and collagen I which both lack a heparin binding domain it was found that multilayers could bind pFN specifically because only this protein was capable of promoting cell adhesion. Multilayer surfaces that inhibited MG-63 adhesion did also cause a decreased cell growth in the presence of serum, while an enhanced adhesion of cells was connected to an improved cell growth.

Keywords: Cell-adhesion, Polyelectrolyte multilayers, Substratum chemistry, Surface-properties, Fibroblast-growth, Fibronectin, Polymers, Chitosan, Polysaccharides, Wettability

Caballero, D., Samitier, J., Errachid, A., (2009). Submerged nanocontact printing (SnCP) of thiols Journal of Nanoscience and Nanotechnology , 9, (11), 6478-6482

Biological patterned surfaces having sub-micron scale resolution are of great importance in many fields of life science and biomedicine. Different techniques have been proposed for surface patterning at the nanoscale. However, most of them present some limitations regarding the patterned area size or are time-consuming. Micro/nanocontact printing is the most representative soft lithography-based technique for surface patterning at the nanoscale. Unfortunately, conventional micro/nanocontact printing also suffers from problems such as diffusion and stamp collapsing that limit pattern resolution. To overcome these problems, a simple way of patterning thiols under liquid media using submerged nanocontact printing (SnCP) over large areas (similar to cm(2)) achieving nanosize resolution is presented. The technique is also low cost and any special equipment neither laboratory conditions are required. Nanostructured poly(dimethyl siloxane) stamps are replicated from commercially available digital video disks. SnCP is used to stamp patterns of 200 nm 1-octadecanethiol lines in liquid media, avoiding ink diffusion and stamp collapsing, over large areas on gold substrates compared with conventional procedures. Atomic force microscopy measurements reveal that the patterns have been successfully transferred with high fidelity. This is an easy, direct, effective and low cost methodology for molecule patterning immobilization which is of interest in those areas that require nanoscale structures over large areas, such as tissue engineering or biosensor applications.

Keywords: Submerged Nanocontact Printing, Replica Molding, Nanopatterning, Large Area, Dip-pen nanolithography, High-aspect-ratio, Soft lithography, Submicronscale, Nanoimprint lithography, Thin-film, Surfaces, Fabrication, Proteins, Nanofabrication

Rodriguez-Segui, S. A., Bucior, I., Burger, M. M., Errachid, A., Fernàndez-Busquets, X., (2009). Application of the quartz crystal microbalance to the study of multivalent carbohydrate-carbohydrate adhesion Sensor Letters 6th Maghreb-Europe Meeting on Materials and Their Applications for Devices and Physical, Chemical and Biological Sensors , AMER SCIENTIFIC PUBLISHERS (Rabat, Morocco) 7, (5), 782-787

Carbohydrate-carbohydrate interactions in cell adhesion are being increasingly explored as important players in cell-cell and cell-extracellular matrix interactions that are characterized by finelytuned on-off rates. The emerging field of glycomics requires the application of new methodologies to the study of the generally weak and multivalent carbohydrate binding sites. Here we use the quartz crystal microbalance (QCM) for the analysis of the self-binding activity of the g200 glycan, a molecule of marine sponge origin that is responsible for Ca2+-dependent species-specific cell adhesion. The QCM has the advantages over other highly sensitive techniques of having only one of the interacting partners bound to a surface, and of lacking microfluidics circuits prone to be clogged by self-aggregating glycans. Our results show that g200 self-interaction is negligible in the absence of Ca2+. Different association kinetics at low and high Ca2+ concentrations suggest the existence of two different Ca2+ binding sites in g200. Finally, the observation of a non-saturable binding indicates that g200 has more than one self-adhesion site per molecule. This work represents the first report to date using the QCM to study carbohydrate-carbohydrate interactions involved in cell adhesion.

Keywords: Ca2+-dependent binding, Carbohydrate-carbohydrate interaction, Cell adhesion, Proteoglycan, Quartz crystal microbalance, Sponges

Planell, J. A., Navarro, M., (2009). Challenges in bone repair Bone repair biomaterials (ed. Planell, J. A., Lacroix, D., Best, S., Merolli, A.), Woodhead (Cambridge, UK) , 3-24

A fundamental aspect of the rapidly expanding medical care sector, bone repair continues to benefit from emerging technological developments. This text provides researchers and students with a comprehensive review of the materials science and engineering principles behind these developments. The first part reviews the fundamentals of bone repair and regeneration. Further chapters discuss the science and properties of biomaterials used in bone repair, including both metals and biocomposites. Final chapters analyze device considerations such as implant lifetime and failure, and discuss potential applications, as well as the ethical issues that continually confront researchers and clinicians.

Keywords: Social impact of musculoskeletal disease, Economic burden of musculoskeletal disease, Social aspects of dental and maxillofacial conditions, Some clinical challenges of bone repair, Conclusions and future trends, Sources of further information and advice

Mateos-Timoneda, M. A., (2009). Polymers for bone repair Bone repair biomaterials (ed. Planell, J. A., Lacroix, D., Best, S., Merolli, A.), Woodhead (Cambridge, UK) , 3-24

A fundamental aspect of the rapidly expanding medical care sector, bone repair continues to benefit from emerging technological developments. This text provides researchers and students with a comprehensive review of the materials science and engineering principles behind these developments. The first part reviews the fundamentals of bone repair and regeneration. Further chapters discuss the science and properties of biomaterials used in bone repair, including both metals and biocomposites. Final chapters analyze device considerations such as implant lifetime and failure, and discuss potential applications, as well as the ethical issues that continually confront researchers and clinicians.

Keywords: Ultra high molecular weight polyethylene (UHMWPE), Acrylic polymers as bone cement, Biodegradable polymers

Engel, E., Del Valle, S., Aparicio, C., Altankov, G., Asin, L., Planell, J. A., Ginebra, M. P., (2008). Discerning the role of topography and ion exchange in cell response of bioactive tissue engineering scaffolds Tissue Engineering Part A , 14, (8), 1341-1351

Surface topography is known to have an influence on osteoblast activity. However, in the case of bioactive materials, topographical changes can affect also ion exchange properties. This makes the problem more complex, since it is often difficult to separate the strictly topographical effects from the effects of ionic fluctuations in the medium. The scope of this paper is to analyze the simultaneous effect of topography and topography-mediated ion exchange on the initial cellular behavior of osteoblastic-like cells cultured on bioactive tissue engineering substrates. Two apatitic substrates with identical chemical composition but different micro/nanostructural features were obtained by low-temperature setting of a calcium phosphate cement. MG63 osteoblastic-like cells were cultured either in direct contact with the substrates or with their extracts. A strong and permanent decrease of calcium concentration in the culture medium, dependent on substrate topography, was detected. A major effect of the substrate microstructure on cell proliferation was observed, explained in part by the topography-mediated ion exchange, but not specifically by the ionic Ca(2+) fluctuations. Cell differentiation was strongly enhanced when cells were cultured on the finer substrate. This effect was not explained by the chemical modification of the medium, but rather suggested a strictly topographical effect.

Keywords: Alkaline Phosphatase/metabolism, Bone Cements/pharmacology, Calcium/metabolism, Calcium Phosphates/pharmacology, Cell Adhesion/drug effects, Cell Differentiation/drug effects, Cell Proliferation/drug effects, Cell Shape/drug effects, Cells, Cultured, Culture Media, Durapatite/pharmacology, Humans, Interferometry, Ion Exchange, Materials Testing, Osteoblasts/ cytology/drug effects/enzymology/ultrastructure, Phosphorus/metabolism, Powders, Tissue Engineering, Tissue Scaffolds

Olmedo, Ivonne, Araya, Eyleen, Sanz, Fausto, Medina, Elias, Arbiol, Jordi, Toledo, Pedro, Àlvarez-Lueje, Alejandro, Giralt, Ernest, Kogan, Marcelo J., (2008). How changes in the sequence of the peptide CLPFFD-NH2 can modify the conjugation and stability of gold nanoparticles and their affinity for beta-amyloid fibrils Bioconjugate Chemistry , 19, (6), 1154-1163

In a previous work, we studied the interaction of β-amyloid fibrils (Aβ) with gold nanoparticles (AuNP) conjugated with the peptide CLPFFD-NH2. Here, we studied the effect of changing the residue sequence of the peptide CLPFFD-NH2 on the efficiency of conjugation to AuNP, the stability of the conjugates, and the affinity of the conjugates to the Aβ fibrils. We conjugated the AuNP with CLPFFD-NH2 isomeric peptides (CDLPFF-NH2 and CLPDFF-NH2) and characterized the resulting conjugates with different techniques including UV−Vis, TEM, EELS, XPS, analysis of amino acids, agarose gel electrophoresis, and CD. In addition, we determined the proportion of AuNP bonded to the Aβ fibrils by ICP-MS. AuNP-CLPFFD-NH2 was the most stable of the conjugates and presented more affinity for Aβ fibrils with respect to the other conjugates and bare AuNP. These findings help to better understand the way peptide sequences affect conjugation and stability of AuNP and their interaction with Aβ fibrils. The peptide sequence, the steric effects, and the charge and disposition of hydrophilic and hydrophobic residues are crucial parameters when considering the design of AuNP peptide conjugates for biomedical applications.

Keywords: Self-assembled monolayers, Aggregation, Dispersions, Adsorption, Particles, Design, Size

Navarro, M., Benetti, E. M., Zapotoczny, S., Planell, J. A., Vancso, G. J., (2008). Buried, covalently attached RGD peptide motifs in poly(methacrylic acid) brush layers: The effect of brush structure on cell adhesion Langmuir 24, (19), 10996-11002

Iniferter-mediated surface-initiated photopolymerization was used to graft poly(methacrylic acid) (PMAA) brush layers obtained from surface-attached iniferters in self-assembled monolayers to a gold surface. The tethered chains were subsequently functionalized with the cell-adhesive arginine-glycine-aspartic acid (RGD) motif. The modified brushes were extended by reinitiating the polymerization to obtain an additional layer of PMAA, thereby burying the peptide-functionalized segments inside the brush structure. Contact angle measurements and Fourier transform infrared (FTIR) spectroscopy were employed to characterize the wettability and the chemical properties of these platforms. Time of flight secondary ion mass spectroscopy (TOF-SIMS) measurements were performed to monitor the chemical composition of the polymer layer as a function of the distance to the gold surface and obtain information concerning the depth of the RGD motifs inside the brush structure. The brush thickness was evaluated as a function of the polymerization (i.e.. UV-irradiation) time with atomic force microscopy (AFM) and ellipsometry. Cell adhesion tests employing human osteoblasts were performed on substrates with the RGD peptides exposed at the surface as well as covered by a PMAA top brush layer. Immunofluorescence studies demonstrated a variation of the cell morphology as a function of the position of the peptide units along the grafted chains.

Keywords: Ion mass-spectrometry, Transfer radical polymerization, Asymmetric diblock copolymers, Arg-gly-asp, Swelling behaviour, Endothelial-cells, Thin-films, fibronectin, Surfaces, SIMS

Navarro, M., Michiardi, A., Castano, O., Planell, J. A., (2008). Biomaterials in orthopaedics Journal of the Royal Society Interface , 5, (27), 1137-1158

At present, strong requirements in orthopaedics are still to be met, both in bone and joint substitution and in the repair and regeneration of bone defects. In this framework, tremendous advances in the biomaterials field have been made in the last 50 years where materials intended for biomedical purposes have evolved through three different generations, namely first generation (bioinert materials), second generation (bioactive and biodegradable materials) and third generation (materials designed to stimulate specific responses at the molecular level). In this review, the evolution of different metals, ceramics and polymers most commonly used in orthopaedic applications is discussed, as well as the different approaches used to fulfil the challenges faced by this medical field.

Keywords: Biomaterials, Orthopaedics, Tissue engineering, Bioactive materials, Biodegradable materials, Bioinert materials

Oncins, Gerard, Vericat, Carolina, Sanz, Fausto, (2008). Mechanical properties of alkanethiol monolayers studied by force spectroscopy Journal of Chemical Physics , 128, (4), 044701

The mechanical properties of alkanethiol monolayers on Au(111) in KOH solution have been studied by force spectroscopy. The analysis of the vertical force versus penetration curves showed that monolayer penetration is a stepped process that combines elastic regions with sudden penetration events. The structural meaning of these events can be explained both by the creation of gauche defects on the hydrocarbon chains and by a cooperative molecular tilting model proposed by Barrena et al. [J. Chem. Phys. 113, 2413 (2000)]. The validity of these models for alkanethiol monolayers of different compactness and chain length has been discussed. The Young's modulus (E) of the monolayers has been calculated by using a recently developed model which considers the thickness of the monolayer as a parameter, thus allowing a decoupling of the mechanical properties of the thiol layer from those of the Au(111) substrate. As a result, the calculated E values are in the range of 50-150 Pa, which are remarkably lower than those previously reported in the literature.

Keywords: Adsorbed layers, AFM, Gold, Monolayers, Organic compounds, Self-assemblyYoung's modulus

Maneva-Radicheva, L., Ebert, U., Dimoudis, N., Altankov, G., (2008). Fibroblast remodeling of adsorbed collagen type IV is altered in contact with cancer cells Histology and Histopathology , 23, (7), 833-842

A series of co-culture experiments between fibroblasts and H-460 human lung carcinoma cells were performed to learn more about the fate of adsorbed type IV collagen (Coll IV). Fibroblasts were able to spatially rearrange Coll IV in a specific linear pattern, similar but not identical to the fibronectin (FN) fibrils. Coll IV partly co-aligns with fibroblast actin cytoskeleton and transiently co-localize with FN, as well as with beta 1 and a 2 integrin clusters, suggesting a cell-dependent process. We further found that this Coll IV reorganization is suppressed in contact with H460 cells. Zymography revealed strongly elevated MMP-2 activity in supernatants of co-cultures, but no activity when fibroblasts or cancer cells were cultured alone. Thus, we provide evidence that reorganization of substrate associated Coll IV is a useful morphological approach for in vitro studies on matrix remodeling activity during tumorigenesis.

Keywords: Adsorbed collagen IV reorganization, Fibroblasts and cancer cells co-culture, MMP-2

Martinez, E., Engel, E., Lopez-Iglesias, C., Mills, C. A., Planell, J. A., Samitier, J., (2008). Focused ion beam/scanning electron microscopy characterization of cell behavior on polymer micro-/nanopatterned substrates: A study of cell-substrate interactions Micron , 39, (2), 111-116

Topographic micro and nanostructures can play an interesting role in cell behaviour when cells are cultured on these kinds of patterned substrates. It is especially relevant to investigate the influence of the nanometric dimensions topographic features on cell morphology, proliferation, migration and differentiation. To this end, some of the most recent fabrication technologies, developed for the microelectronics industry, can be used to produce well-defined micro and nanopatterns on biocompatible polymer substrates. In this work, osteoblast-like cells are grown on poly(methyl methacrylate) substrates patterned by nanoimprint lithography techniques. Examination of the cell-substrate interface can reveal important details about the cell morphology and the distribution of the focal contacts on the substrate surface. For this purpose, a combination of focused ion beam milling and scanning electron microscopy techniques has been used to image the cell-substrate interface. This technique, if applied to samples prepared by freeze-drying methods, allows high-resolution imaging of cross-sections through the cell and the substrate, where the interactions between the nanopatterned substrate, the cell and the extracellular matrix, which are normally hidden by the bulk of the cell, can be studied.

Keywords: Electron microscopy, Interface, Nanotopography, Osteoblast, Adhesion molecule, Cell morphology

Gustavsson, J., Altankov, G., Errachid, A., Samitier, J., Planell, J. A., Engel, E., (2008). Surface modifications of silicon nitride for cellular biosensor applications Journal of Materials Science-Materials in Medicine , 19, (4), 1839-1850

Thin films of silicon nitride (Si3N4) can be used in several kinds of micro-sized biosensors as a material to monitor fine environmental changes related to the process of bone formation in vitro. We found however that Si3N4 does not provide optimal conditions for osseointegration as osteoblast-like MG-63 cells tend to detach from the surface when cultured over confluence. Therefore Si3N4 was modified with self-assembled monolayers bearing functional end groups of primary amine (NH2) and carboxyl (COOH) respectively. Both these modifications enhanced the interaction with confluent cell layers and thus improve osseointegration over Si3N4. Furthermore it was observed that the NH2 functionality increased the adsorption of fibronectin (FN), promoted cell proliferation, but delayed the differentiation. We also studied the fate of pre-adsorbed and secreted FN from cells to learn more about the impact of above functionalities for the development of provisional extracellular matrix on materials interface. Taken together our data supports that Si3N4 has low tissue integration but good cellular biocompatibility and thus is appropriate in cellular biosensor applications such as the ion-sensitive field effect transistor (ISFET). COOH and NH2 chemistries generally improve the interfacial tissue interaction with the sensor and they are therefore suitable substrates for monitoring cellular growth or matrix deposition using electrical impedance spectroscopy.

Keywords: Adsorption, Amines/chemistry, Biocompatible Materials/ chemistry, Biosensing Techniques, Cell Differentiation, Cell Line, Cell Proliferation, Electric Impedance, Extracellular Matrix/metabolism, Fibronectins/chemistry, Humans, Materials Testing, Osteoblasts/ cytology, Silicon Compounds/ chemistry, Surface Properties

Diez, Pablo F., Laciar, Eric, Mut, Vicente, Avila, Enrique, Torres, Abel, (2008). A comparative study of the performance of different spectral estimation methods for classification of mental tasks IEEE Engineering in Medicine and Biology Society Conference Proceedings 30th Annual International Conference of the Ieee Engineering in Medicine and Biology Society (ed. IEEE), IEEE (Vancouver, Canada) 1-8, 1155-1158

In this paper we compare three different spectral estimation techniques for the classification of mental tasks. These techniques are the standard periodogram, the Welch periodogram and the Burg method, applied to electroencephalographic (EEG) signals. For each one of these methods we compute two parameters: the mean power and the root mean square (RMS), in various frequency bands. The classification of the mental tasks was conducted with a linear discriminate analysis. The Welch periodogram and the Burg method performed better than the standard periodogram. The use of the RMS allows better classification accuracy than the obtained with the power of EEG signals.

Keywords: Adult, Algorithms, Artificial Intelligence, Cognition, Electroencephalography, Female, Humans, Male, Pattern Recognition, Automated, Reproducibility of Results, Sensitivity and Specificity, Task Performance and Analysis, User-Computer Interface

Casuso, I., Pla, M., Gomila, G., Samitier, J., Minic, J., Persuy, M. A., Salesse, R., Pajot-Augy, E., (2008). Immobilization of olfactory receptors onto gold electrodes for electrical biosensor Materials Science & Engineering C 5th Maghreb-Europe Meeting on Materials and their Applicatons for Devices and Physical, Chemical and Biological Sensors , Elsevier Science (Mahdia, TUNISIA) 28, (5-6), 686-691

We investigate the immobilization of native nanovesicles containing functional olfactory receptors onto gold electrodes by means of atomic force microscopy in liquid. We show that nanovesicles can be adsorbed without disrupting them presenting sizes once immobilized ranging from 50 run to 200 nm in diameter. The size of the nanovesicles shows no dependence on the electrode hydrophobicity being constant in a height/width ratio close to 1:3. Nevertheless, electrode hydrophobicity does affect the surface coverage, the surface coverage is five times higher in hydrophilic electrodes than on hydrophobic ones. Surface coverage is also affected by nanovesicles dimensions in suspension, the size homogenization to around 50 nm yields a further five fold increment in surface coverage achieving a coverage of about 50% close to the hard spheres jamming limit (54.7%). A single layer of nanovesicles is always formed with no particle overlap. Present results provide insights into the immobilization on electrodes of olfactory receptors for further olfactory electrical biosensor development.

Keywords: AFM, Adsorption, Odorant, Taste

Hernando, Jordi, Hoogenboom, Jacob, van Dijk, Erik, Garcia-Parajo, Maria, van Hulst, Niek F., (2008). Ultrafast single-molecule photonics: Excited state dynamics in coherently coupled complexes Journal of Luminescence 16th International Conference on Dynamical Processes in Excited States of Solids (ed. -----), Elsevier Science BV (Segovia, Spain) 128, (5-6), 1050-1052

We present a single-molecule study on femtosecond dynamics in multichromophoric systems, combining fs pump-probe, emission-spectra and fluorescence-lifetime analysis. The ultrafast fs approach gives direct information on the initial exciton dynamics after excitation. The lifetime data show superradiance, a direct measure for the extent of the coherent coupling and static disorder. The spectra finally reveal the role of exciton-phonon coupling. At the single-molecule level a wide range of exciton delocalization lengths and energy redistribution times is revealed.

Keywords: Single-molecule detection, Pump-probe, Exciton delocalization, Superradiance, Exciton-phonon coupling

Charles-Harris, M., del Valle, S., Hentges, E., Bleuet, P., Lacroix, D., Planell, J. A., (2007). Mechanical and structural characterisation of completely degradable polylactic acid/calcium phosphate glass scaffolds Biomaterials 28, (30), 4429-4438

This study involves the mechanical and structural characterisation of completely degradable scaffolds for tissue engineering applications. The scaffolds are a composite of polylactic acid (PLA) and a soluble calcium phosphate glass, and are thus completely degradable. A factorial experimental design was applied to optimise scaffold composition prior to simultaneous microtomography and micromechanical testing. Synchrotron X-ray microtomography combined with in situ micromechanical testing was performed to obtain three-dimensional 3D images of the scaffolds under compression. The 3D reconstruction was converted into a finite element mesh which was validated by simulating a compression test and comparing it with experimental results. The experimental design reveals that larger glass particle and pore sizes reduce the stiffness of the scaffolds, and that the porosity is largely unaffected by changes in pore sizes or glass weight content. The porosity ranges between 93% and 96.5%, and the stiffness ranges between 50 and 200 kPa. X-ray projections show a homogeneous distribution of the glass particles within the PLA matrix, and illustrate pore-wall breakage under strain. The 3D reconstructions are used qualitatively to visualise the distribution of the phases of the composite material, and to follow pore deformation under compression. Quantitatively, scaffold porosity, pore interconnectivity and surface/volume ratios have been calculated. Finite element analysis revealed the stress and strain distribution in the scaffold under compression, and could be used in the future to characterise the mechanical properties of the scaffolds.

Keywords: Synchrotron x-ray microtomography, Mechanical test, Biodegradable, Glass, Scaffold, Finite element analysis

Pla, M., Fernandez, Javier G., Mills, C. A., Martinez, E., Samitier, J., (2007). Micro/nanopatterning of proteins via contact printing using high aspect ratio PMMA stamps and NanoImprint apparatus Langmuir 23, (16), 8614-8618

Micro- and nanoscale protein patterns have been produced via a new contact printing method using a nanoimprint lithography apparatus. The main novelty of the technique is the use of poly(methyl methacrylate) (PMMA) instead of the commonly used poly(dimethylsiloxane) (PDMS) stamps. This avoids printing problems due to roof collapse, which limits the usable aspect ratio in microcontact printing to 10:1. The rigidity of the PMMA allows protein patterning using stamps with very high aspect ratios, up to 300 in this case. Conformal contact between the stamp and the substrate is achieved because of the homogeneous pressure applied via the nanoimprint lithography instrument, and it has allowed us to print lines of protein similar to 150 nm wide, at a 400 nm period. This technique, therefore, provides an excellent method for the direct printing of high-density sub-micrometer scale patterns, or, alternatively, micro-/nanopatterns spaced at large distances. The controlled production of these protein patterns is a key factor in biomedical applications such as cell-surface interaction experiments and tissue engineering.

Keywords: Soft lithography, Cell-adhesion, Microstructures, Fabrication, Stability, Patterns

Rico, Félix, Roca-Cusachs, Pere, Sunyer, Raimon, Farré, Ramon, Navajas, Daniel, (2007). Cell dynamic adhesion and elastic properties probed with cylindrical atomic force microscopy cantilever tips Journal of Molecular Recognition John Wiley & Sons, Ltd. 20, (6), 459-466

Cell adhesion is required for essential biological functions such as migration, tissue formation and wound healing, and it is mediated by individual molecules that bind specifically to ligands on other cells or on the extracellular matrix. Atomic force microscopy (AFM) has been successfully used to measure cell adhesion at both single molecule and whole cell levels. However, the measurement of inherent cell adhesion properties requires a constant cell-probe contact area during indentation, a requirement which is not fulfilled in common pyramidal or spherical AFM tips. We developed a procedure using focused ion beam (FIB) technology by which we modified silicon pyramidal AFM cantilever tips to obtain flat-ended cylindrical tips with a constant and known area of contact. The tips were validated on elastic gels and living cells. Cylindrical tips showed a fairly linear force-indentation behaviour on both gels and cells for indentations > 200nm. Cylindrical tips coated with ligands were used to quantify inherent dynamic cell adhesion and elastic properties. Force, work of adhesion and elasticity showed a marked dynamic response. In contrast, the deformation applied to the cells before rupture was fairly constant within the probed dynamic range. Taken together, these results suggest that the dynamic adhesion strength is counterbalanced by the dynamic elastic response to keep a constant cell deformation regardless of the applied pulling rate.

Keywords: AFM, Cell adhesion, Cell mechanics, Cell stiffness

Rodriguez, Segui, Bucior, I., Burger, M. M., Samitier, J., Errachid, A., Fernàndez-Busquets, X., (2007). Application of a bio-QCM to study carbohydrates self-interaction in presence of calcium Transducers '07 & Eurosensors Xxi, Digest of Technical Papers 14th International Conference on Solid-State Sensors, Actuators and Microsystems , IEEE (Lyon, France) 1-2, 1995-1998

In the past years, the quartz crystal microbalance (QCM) has been successfully applied to follow interfacial physical chemistry phenomena in a label free and real time manner. However, carbohydrate self adhesion has only been addressed partially using this technique. Carbohydrates play an important role in cell adhesion, providing a highly versatile form of attachment, suitable for biologically relevant recognition events in the initial steps of adhesion. Here, we provide a QCM study of carbohydrates' self-recognition in the presence of calcium, based on a species-specific cell recognition model provided by marine sponges. Our results show a difference in adhesion kinetics when varying either the calcium concentration (with a constant carbohydrate concentration) or the carbohydrate concentration (with constant calcium concentration).

Keywords: Biomedical materials, Calcium, Cellular biophysics, Microbalances, Porous materials, Quartz, Surface chemistry/ bio-QCM, Carbohydrates self-interaction, Quartz crystal microbalance, Interfacial physical chemistry phenomena, Carbohydrate self adhesion, Biologically relevant recognition events, Marine sponges, Adhesion kinetics, Calcium concentration, Carbohydrate concentration, Biosensors, Biomedical materials, Surface chemistry, Cellular biophysics