Newly Engineered Magnetic Erythrocytes for Sustained and Targeted Delivery of Anti-Cancer Therapeutic Compounds

Cytotoxic chemotherapy of cancer is limited by serious, sometimes life-threatening, side effects that arise from toxicities to sensitive normal cells because the therapies are not selective for malignant cells. So how can they be selectively improved? Alternative pharmaceutical formulations of anti-cancer agents have been investigated in order to improve conventional chemotherapy treatment. These formulations are associated with problems like severe toxic side effects on healthy organs, drug resistance and limited access of the drug to the tumor sites suggested the need to focus on site-specific controlled drug delivery systems. In response to these concerns, we have developed a new drug delivery system based on magnetic erythrocytes engineered with a viral spike fusion protein. This new erythrocyte-based drug delivery system has the potential for magnetic-controlled site-specific localization and highly efficient fusion capability with the targeted cells. Here we show that the erythro-magneto-HA virosomes drug delivery system is able to attach and fuse with the target cells and to efficiently release therapeutic compounds inside the cells. The efficacy of the anti-cancer drug employed is increased and the dose required is 10 time less than that needed with conventional therapy

Sleep-related hypermotor epilepsy: Long-term outcome in a large cohort

To assess the long-term outcome of sleep-related hypermotor epilepsy (SHE)

Neuroradiological study (morphological and functional) in patients with Nocturnal Frontal Lobe Epilepsy

L'epilessia frontale notturna (EFN) è caratterizzata da crisi motorie che insorgono durante il sonno. Scopo del progetto è studiare le cause fisiopatologiche e morfo-funzionali che sottendono ai fenomeni motori nei pazienti con EFN e identificare alterazioni strutturali e/o metaboliche mediante tecniche avanzate di Risonanza Magnetica (RM). Abbiamo raccolto una casistica di pazienti con EFN afferenti al Centro Epilessia e dei Disturbi del Sonno del Dipartimento di Scienze Neurologiche, Università di Bologna. Ad ogni paziente è stato associato un controllo sano di età (± 5 anni) e sesso corrispondente. Tutti sono stati studiati mediante tecniche avanzate di RM comprendenti Spettroscopia del protone (1H-MRS), Tensore di diffusione ed imaging 3D ad alta risoluzione per analisi morfometriche. In particolare, la 1H-MRS è stata effettuata su due volumi di interesse localizzati nei talami e nel giro del cingolo anteriore. Sono stati inclusi nell’analisi finale 19 pazienti (7 M), età media 34 anni (range 19-50) e 14 controlli (6 M) età media 30 anni (range 19-40). A livello del cingolo anteriore il rapporto della concentrazione di N-Acetil-Aspartato rispetto alla Creatina (NAA/Cr) è risultato significativamente ridotto nei pazienti rispetto ai controlli (p=0,021). Relativamente all’analisi di correlazione, l'analisi tramite modelli di regressione multipla ha evidenziato che il rapporto NAA/Cr nel cingolo anteriore nei pazienti correlava con la frequenza delle crisi (p=0,048), essendo minore nei pazienti con crisi plurisettimanali/plurigiornaliere. Per interpretare il dato ottenuto è possibile solo fare delle ipotesi. L’NAA è un marker di integrità, densità e funzionalità neuronale. E’ possibile che alla base della EFN ci siano alterazioni metaboliche tessutali in precise strutture come il giro del cingolo anteriore. Questo apre nuove possibilità sull’utilizzo di strumenti di indagine basati sull’analisi di biosegnali, per caratterizzare aree coinvolte nella genesi della EFN ancora largamente sconosciute e chiarire ulteriormente l’eziologia di questo tipo di epilessia.Nocturnal frontal lobe epilepsy (NFLE) is characterized by motor seizuresoccurring during sleep. Aims of the study were to identify the pathophysiological and anatomo-functional factors implicated in the genesis of the motor episodes and possible structural alterations and/or metabolic disorders ,in NFLE patients using advanced techniques of Magnetic Resonance (MR). We have collected a series of patients with NFLE attending the Epilepsy and Sleep Centres of the Department of Neurological Sciences, Bologna University. For each NFLE patient, one control subject of the same sex and age (±5 years) was recruited. All subjects were studied using proton spectroscopy (1H-MRS), diffusion tensor imaging and high resolution 3D-imaging for morphometric analyses. In particular, 1H-MRS was performed in two regions of interest: the thalami and the anterior cingulate gyrus. Nineteen patients (7M), mean age 34 years (range 19-50) and 14 controls (6 M), mean age 30 years (range 19-40), were included in the final analysis. At the level of the anterior cingulate the ratio of the concentration of N-Acetyl-Aspartate compared to Creatine (NAA/Cr) was significantly reduced in patients compared with controls (p=0,021). A multiple regression analysis was performed, showing that the NAA/Cr ratio in the anterior cingulate in patients correlated with the seizures frequency (p=0,048), being lower in patients with very frequent seizures. As NAA is a marker of integrity, density and neuronal function, we speculate that metabolic changes in specific structures, such as the anterior cingulate, may underlie the pathogenesis of NFLE. Our results open up new possibilities for the use of methodologies of biomedical signal/image processing to characterize the areas involved in the genesis of NFLE, which are currently largely unknown and to further clarify the etiology of this type of epilepsy

Discontinuous Galerkin Approximation of Relaxation Models for Linear and Nonlinear Diffusion Equations

In this work we present finite element approximations of relaxed systems for nonlinear diffusion problems, which can also tackle the cases of degenerate and strongly degenerate diffusion equations. Relaxation schemes take advantage of the replacement of the original partial differential equation (PDE) with a semilinear hyperbolic system of equations, with a stiff source term, tuned by a relaxation parameter \u3b5. When \u3b5 \u2192 0+, the system relaxes onto the original PDE: in this way, a consistent discretization of the relaxation system for vanishing \u3b5 yields a consistent discretization of the original PDE. The numerical schemes obtained with this procedure do not require solving implicit nonlinear problems and possess the robustness of upwind discretizations. The proposed approximations are based on a discontinuous Galerkin method in space and on suitable implicitexplicit integration in time. Then, in principle, we can achieve any order of accuracy and obtain stable solutions, even when the diffusion equation becomes degenerate and solution singularities develop. Moreover, when needed, we can easily incorporate slope limiters within our schemes in order to handle spurious oscillatory phenomena. Some preliminary theoretical results are given, along with several numerical tests in one and two space dimensions, both for linear and nonlinear diffusion problems, including a degenerate diffusion equation, that provide numerical evidence of the properties of the presented approach

FACS analysis of HeLa cells after (A) 24 h, (B) 48 h and (C) 96 h treatments.

<p><b>Ctrl</b> (control) untreated cells; <b>Aza:</b> cells treated with 1800 ng (2,5 µM) of 5-Aza-2-dC; <b>Erythro Aza:</b> cells treated with erythro-magneto-HA virosomes containing 180 ng of 5-Aza-2-dC; <b>Erythro:</b> unloaded 5-Aza-2-dC erythro-magneto-HA virosomes; <b>Supernatant:</b> cells treated with buffer where the erythro-magneto-HA virosomes were resuspended (control 2).</p

Confocal Laser Scanning Microscopy (CLSM).

<p>In the left is schematized the timing and mechanism of action of the erythro-magneto-HA virosomes, encapsulating 100 nm fluorescence-labelled magnetic nanoparticles (green) and 5-Aza-2-dC. In the right are shown CLSM images of erythro-magneto-HA virosomes (green) after 30 minutes (<b>A</b>), 1 hour (<b>B</b>), 6 hours (<b>C</b>), 24 and 96 hours (<b>D</b>) of incubation with HeLa cells highlighted by DAPI staining (blue). (<b>CTRL</b>) Untreated HeLa cells (control).</p

Engineered Erythrocytes.

<p>Confocal Laser Scanning Microscopy (CLSM) images of erytro-magneto-HA virosomes encapsulating 100 nm fluorescence-labelled superparamagnetic nanoparticles (green) and 5-Aza-2-dC.</p