The type of interaction between tumor-associated antigens and specialized antigen-presenting cells such as dendritic cells (DCs) is critical for the type of immunity that will be generated. MUC1, a highly O-glycosylated mucin, is overexpressed and aberrantly glycosylated in several tumor histotypes. This results in the expression of tumor-associated glycoforms and in MUC1 carrying the tumor-specific glycan Tn (GalNAcα1-O-Ser/Thr). Glycopeptides corresponding to three tandem repeats of MUC1, enzymatically glycosylated with 9 or 15 mol of GalNAc, were shown to specifically bind and to be internalized by immature monocyte-derived DCs (iDCs). Binding required calcium and the GalNAc residue and was competed out by GalNAc polymer and Tn-MUC1 or Tn-MUC2 glycopeptides. The macrophage galactose-type C-type lectin (MGL) receptor expressed on iDCs was shown to be responsible for the binding. Confocal analysis and ELISA done on subcellular fractions of iDCs showed that the Tn-MUC1 glycopeptides colocalized with HLA class I and II compartments after internalization. Importantly, although Tn-MUC1 recombinant protein was bound and internalized by MGL, the glycoprotein entered the HLA class II compartment, but not the HLA class I pathway. These data indicate that MGL expressed on iDCs is an optimal receptor for the internalization of short GalNAcs carrying immunogens to be delivered into HLA class I and II compartments. Such glycopeptides therefore represent a new way of targeting the HLA class I and II pathways of DCs. These results have possible implications in designing cancer vaccines. [Cancer Res 2007;67(17):8358–67]

Mutations in the PINK1 gene cause autosomal recessive Parkinson's disease. The PINK1 gene encodes a protein kinase that is mitochondrially cleaved to generate two mature isoforms. In addition to its protective role against mitochondrial dysfunction and apoptosis, PINK1 is also known to regulate mitochondrial dynamics acting upstream of the PD-related protein Parkin. Recent data showed that mitochondrial Parkin promotes the autophagic degradation of dysfunctional mitochondria, and that stable PINK1 silencing may have an indirect role in mitophagy activation. Here we report a new interaction between PINK1 and Beclin1, a key pro-autophagic protein already implicated in the pathogenesis of Alzheimer's and Huntington's diseases. Both PINK1 N- and C-terminal are required for the interaction, suggesting that full-length PINK1, and not its cleaved isoforms, interacts with Beclin1. We also demonstrate that PINK1 significantly enhances basal and starvation-induced autophagy, which is reduced by knocking down Beclin1 expression or by inhibiting the Beclin1 partner Vps34. A mutant, PINK1^W437X, interaction of which with Beclin1 is largely impaired, lacks the ability to enhance autophagy, whereas this is not observed for PINK1^G309D, a mutant with defective kinase activity but unaltered ability to bind Beclin1. These findings identify a new function of PINK1 and further strengthen the link between autophagy and proteins implicated in the neurodegenerative process.

The aim of this work was to identify micro‐RNAs (miRNAs) involved in the pathological pathways activated in skeletal muscle damage and regeneration by both dystrophin absence and acute ischemia. Eleven miRNAs were deregulated both in MDX mice and in Duchenne muscular dystrophy patients (DMD signature). Therapeutic interventions ameliorating the mdx ‐phenotype rescued DMD‐signature alterations. The significance of DMD‐signature changes was characterized using a damage/regeneration mouse model of hind‐limb ischemia and newborn mice. According to their expression, DMD‐signature miRNAs were divided into 3 classes. 1 ) Regeneration miRNAs, miR‐31, miR‐34c, miR‐206, miR‐335, miR‐449, and miR‐494, which were induced in MDX mice and in DMD patients, but also in newborn mice and in newly formed myofibers during postischemic regeneration. Notably, miR‐206, miR‐34c, and miR‐335 were up‐regulated following myoblast differentiation in vitro. 2 ) Degenerative‐miRNAs, miR‐1, miR‐29c, and miR‐135a, that were down‐modulated in MDX mice, in DMD patients, in the degenerative phase of the ischemia response, and in newborn mice. Their down‐modulation was linked to myofiber loss and fibrosis. 3 ) Inflammatory miRNAs, miR‐222 and miR‐223, which were expressed in damaged muscle areas, and their expression correlated with the presence of infiltrating inflammatory cells. These findings show an important role of miRNAs in physiopathological pathways regulating muscle response to damage and regeneration.—Greco, S., De Simone, M., Colussi, C., Zaccagnini, G., Fasanaro, P., Pescatori, M., Cardani, R., Perbellini, R., Isaia, E., Sale, P., Meola, G., Capogrossi, M. C., Gaetano, C., Martelli, F. Common micro‐RNA signature in skeletal muscle damage and regeneration induced by Duchenne muscular dystrophy and acute ischemia. FASEB J . 23, 3335–3346 (2009). www.fasebj.org

Background— Fabry cardiomyopathy is diagnosed by detection of left ventricular hypertrophy (LVH) in patients with α-Galactosidase A deficiency. Conventional noninvasive tools are unable to provide a preclinical diagnosis allowing prompt institution of enzymatic therapy. Methods and Results— We studied three groups of patients: 10 patients with causal mutations for Fabry disease and LVH, 10 mutation-positive patients without LV, and 10 healthy relatives without causal mutations and no LVH. All patients with LVH and 6 patients with Fabry disease without LVH with complex repetitive ventricular arrhythmias underwent biventricular endomyocardial biopsy to assess cardiac involvement. In all patients 2-dimensional echocardiography with tissue Doppler analysis in the pulsed Doppler mode was performed: systolic (Sa), early diastolic (Ea), and late diastolic (Aa) velocities were measured, and the Ea/Aa ratio and the dimensionless parameter E/Ea were computed at both corners of the mitral annulus. Histology and electron microscopy studies showed glycosphingolipid deposits in all cases. All mutation-positive patients had significant reduction of Sa, Ea, and Aa velocities at both corners of the mitral annulus compared with normal control subjects. Ea/Aa ratio was significantly lower and E/Ea ratio significantly higher in mutation-positive patients than in control subjects. Patients with LVH showed significantly lower contraction and relaxation tissue Doppler velocities, lower Ea/Aa ratio, and higher E/Ea ratio in comparison with mutation-positive patients with no LVH. Conclusions— Fabry cardiomyopathy is characterized by reduced myocardial contraction and relaxation tissue Doppler velocities, detectable even before development of LVH. Tissue Doppler imaging can provide a preclinical diagnosis of Fabry cardiomyopathy, allowing early institution of enzyme replacement therapy.

Objective . A randomized controlled observer-blind trial was designed to evaluate the effectiveness of action observation as an add-on treatment to the standard rehabilitation of upper-limb function, early after stroke. Methods . Stroke survivors (N = 102) were consecutively recruited from 13 centers 30 days (±7) after a first-ever stroke and randomly assigned to the experimental (EG) or control group (CG). EG participants watched video footage of daily routine tasks (actions) carried out with the upper limb in order to prepare to imitate the presented action. At the end of each sequence, a therapist prompted the patient to perform the same movement for 2 minutes, providing help when needed. Static images without animals or human beings were shown to the CG. At the end of each sequence, the CG executed movements that simulated the shoulder and elbow joint mobilization activities performed by the EG. Results for the Fugl-Meyer test, Frenchay Arm test, Box and Block test (BBT), Modified Ashworth Scale, and Functional Independence Measure Motor items were recorded before treatment (T0), after 4 weeks of treatment (T1), and at the follow-up visit 4 to 5 months after the conclusion of treatment (T2). Results . An improvement over time was appreciated on all measures of impairment and functional ability with both treatment programs. A Time × Treatment interaction emerged from the generalized estimating equations analysis of BBT, showing significant T0–T1 and T0–T2 differences in favor of EG. Conclusion . This multicenter trial endorses the use of action observation in upper-extremity rehabilitation, along with a role for the mirror neuron system in poststroke recovery.