Is conscious perception of emotional face expression related to enhanced cortical responses? Electroencephalographic data (112 channels) were recorded in 15 normal adults during the presentation of cue stimuliwith neutral, happy or sad schematic faces (duration: ‘‘threshold time’’ inducing about 50%of correct recognitions), masking stimuli (2 s), and go stimuli with happy or sad schematic faces (0.5 s). The subjects clicked left (right) mouse button in response to go stimuli with happy (sad) faces. After the response, they said ‘‘seen’’ or ‘‘not seen’’ with reference to previous cue stimulus. Electroencephalographic data formed visual event-related potentials (ERPs). Cortical sources of ERPs were estimated by LORETA software. Reaction time to go stimuliwas generally shorter during ‘‘seen’’ than ‘‘not seen’’ trials, possibly due to covert attention and awareness. The cue stimuli evoked four ERP components (posteriorN100, N170, P200, and P300),which had similar peak latency in the ‘‘not seen’’ and ‘‘seen’’ ERPs. Only N170 amplitude showed differences in amplitude in the ‘‘seen’’ versus ‘‘not seen’’ ERPs. Compared to the ‘‘not seen’’ ERPs, the ‘‘seen’’ ones showed prefrontal, premotor, and posterior parietal sources of N170 higher in amplitude with the sad cue stimuli and lower in amplitude with the neutral and happy cue stimuli. These results suggest that nonconscious and conscious processing of schematic emotional facial expressions shares a similar temporal evolution of cortical activity, and conscious processing induces an early enhancement of bilateral cortical activity for the schematic sad facial expressions (N170)

Effects of novel sintetic peroxisome proliferator-activation receptor  (PPARs) agonist on neuronal differentiation in the human neuroblastoma SH-SY5Y cell line MALLAMACI R1, LAGHEZZA A3,LOIODICE F3, VITIELLO F2, BUTTIGLIONE M2 1Dip.Pharmaceutical Biology, Faculty of Pharmacy, University of Bari 2Dip.Biomedical Sciences and Human Oncology, Faculty of Medicine, University of Bari 3Dip. Pharmaceutical Chemistry, Faculty of Pharmacy, University of Bari BACKGROUND: PPARs are a subfamily of the nuclear hormone receptor that heterodimerizes with the retinoid X receptor to act as a transcriptional regulator. Recently it  has been shown that the activation of PPARγ isoform promotes neuronal differentiation. AIM: In this study we have investigated the capability of synthetic compounds endowed with different activity profile on PPARα/γ subtypes to induce neuronal differentiation and neurite outgrowth. The experiments were carried out on cells of  the human neuroblastoma SH-SY5Y line. Retinoic acid (RA) was used as a  positive control for neurite outgrowth. METHODS: SH-SY5Y cultures were maintained in DMEM supplemented with 10% FCS. 10µM RA,  0,5 to 25µM PPAR agonist was added to the culture media in the experimental cultures. Cell viability was tested using the MTT assay. Changes in the expression and re-organization of specific markers involved in neuronal differentiation were investigated using immunofluorescence. RESULTS: Synthetic PPAR agonists promote cell differentiation and the outgrowth of cell processes in a concentration-dependent manner. The maximal effect was obtained at a concentration of 25µM. At this concentration we obtained a significant increase of the expression of neurofilament-200 (NF-H) and Gap-43; this agrees with a hypothesis of cell differentiation induced by these molecules.  CONCLUSION: Our results suggest that sintetic PPAR agonist promotes neuronal differentiation and neurite outgrowth in  SH-SY5Y human neuroblastoma cells. Further work on this subject is underway in our laboratories.

Tourette syndrome (TS) is a chronic multiple-tic disorder affecting 0.8 to 1% of the pediatric population. Consistent data support the presence of abnormalities of immune regulation in children with TS. Environmental factors acting on the immune system, such as group A Streptococcus (GAS) infection, could modify the course of the severity of tics and symptoms in TS. Structural similarity between streptococcal and cerebral antigens might elicit a pathogenic cross-reactivity of antibodies originally targeting GAS antigens to host antigens. Molecular mimicry models has been proposed involving cross-reactivity between streptococcal and neuronal proteins. Given the role of dopamine and dopamine receptors in the control of movement and behavior, we hypothesized that patients with TS harbored serum autoantibodies against dopamine receptors. In our work, we screen and measure the anti-neuronal antibodies in TS patient’s sera. Specifically, we use flow cytometry cell-based assay to test sera for the presence of autoantibodies binding to dopamine D2 (DRD2) and D1 receptors (DRD1) on surface of transfected HEK293 cells. Within the European Multicentre Tics in Children Studies project, financed under the 7th Framework Programme of the European Union, we measure IgG reactivity to DRD2, DRD1, of a thousand of Tourette’s sera from all Europe. The data thus obtained will present high statistical power and will represent the starting point for the diagnostic methodology of TS.

ATP-sensitive-K+ (KATP) channels couple metabolism to the electrical activity of the cells. This channel is associated with glycolytic enzymes to form complexes regulating the channel activity in various tissues. The pyruvate-kinase (PK) enzyme is an antigen in the Paediatric Autoimmune Neuropsychiatric Disorders Associated Streptococcal infection known as PANDAS which is characterized by an abnormal production of auto-antibodies against PK. Here, the effects of the anti-pyruvate kinase antibody (anti-PK-ab) on the muscle and neuronal KATP channels were investigated in native rat skeletal muscle fibres and human neuroblastoma cell-line (SH-SY5Y), respectively. Furthermore, the interaction of PK with the inwardly rectifier potassium channel (Kir6.1/Kir6.2) subunits of the KATP channels was investigated by co-immunoprecipitation experiments in mouse brain using the anti-PK-ab. Patch-clamp experiments showed that the short-term incubation (1 h) of the fibres with the anti-PK-ab at the dilutions of 1:500 and 1:300 enhanced the KATP current of 19.6% and 33.5%, respectively. As opposite, the long-term incubation (24 h) of the fibres with the anti-PK-ab at the dilutions of 1:500 and 1:300 reduced the KATP current of 16% and 24%, respectively, reducing the diameter with atrophy. The direct application of the anti-PK-ab to the excised patches in the absence of intracellular ATP caused channel block, while in the presence of nucleotide channel opened. In neuronal cell line, in the short-term the anti-PK-ab potentiated KATP currents without affecting survival, while in the long-term the anti-PK-ab reduced KATP currents inducing neuronal death. Opening/blocking actions of the anti-PK antibodies on the KATP channels were observed, the blocking action causes fibre atrophy and neuronal death. We demonstrated that PK and Kir subunits are physically/functionally coupled in neurons. The KATP/PK complex can be proposed a novel target in the autoimmune diseases associated with anti-PK production as in PANDAS.

Post-streptococcal neuropsychiatric disorders encompass a broad spectrum of movement disorders, including tics, stereotypies, dystonia and tremor. We report the case of a 15-year-old boy who presented with a relapsing–remitting combination of psychogenic and organic movement disorders. Both relapses occurred after an episode of streptococcal pharyngitis and consisted in motor and phonic tics, an atypical gait disorder, and severe worsening of a pre-existing psychogenic tremor of the right hand. After each relapse, both psychogenic and ‘organic’ symptoms concomitantly remitted after the administration of an association of oral steroids and antibiotics. The peculiarity of this case consists in the coexistence of psychogenic and organic symptoms subsequent to streptococcal infection, and broadens the clinical spectrum of poststreptococcal neuropsychiatric disorders.

It has been shown that concomitant painful stimulation and simple movement at the same hand is related to decreased anticipatory alpha event-related desynchronization (ERD) and reduced pain intensity, possibly due to the interference between somatosensory and motor information processing (Babiloni et al. [6]). Here, we tested the hypothesis that such interference also affects motor performance during sequential movements. Visual warning stimuli were followed by imperative stimuli associated to electrical painful stimulation at left or right middle finger; imperative stimuli triggered motor sequences with right index finger. Electroencephalographic data (N=10, 128 electrodes) were spatially enhanced by surface Laplacian transformation. Cortical activity as revealed by the alpha event-related desynchronization (ERD) was compared in "Pain+ipsilateral movement" condition (movements and painful stimuli performed at the right hand) vs. "Pain+contralateral movement" condition (painful stimuli at left hand and movements performed at the right hand). Results showed that compared with the "Pain+contralateral movement" condition, the "Pain+ipsilateral movement" condition induced lower anticipatory alpha ERD (about 10-12 Hz) in left sensorimotor area, lower subjective pain rate, and delayed movement initiation at the group level. These findings suggest that anticipatory alpha rhythms may underlie cortical preparatory sensorimotor processes preceding somatosensory painful and the initiation of sequential motor events occurring at unilateral or bilateral hand.

AIM To evaluate the relationship between diagnosis and clinical course of Tourette syndrome and group A Streptococcus (GAS). METHOD GAS infections, anti-streptococcal, and anti-basal ganglia antibodies (ABGA) were compared between 168 patients (136males, 32 females) with Tourette syndrome; (median [range] age [25th–75th centile] 10y [8–11y]);median Tourette syndrome duration (25th–75th centile), 3y (1y 3mo–5y 9mo) and a comparison group of 177 patients (117males, 60 females) with epileptic or sleep disorders median age [25th–75th centile], 10y [8y–1y 6mo]). One hundred and forty-four patients with Tourette syndrome were followed up at 3-month intervals; exacerbations of tics, obsessive–compulsive symptoms, and other psychiatric comorbidities were defined by a bootstrap procedure. The effect of new GAS infections and identification of new ABGA upon risk of exacerbation was assessed using logistic regression analysis. RESULTS Cross-sectionally, patients with Tourette syndrome exhibited a higher frequency of GAS infection (8% vs 2%; p=0.009), higher anti-streptolysin O (ASO) titres (246 [108–432] vs 125 [53–269]; p<0.001), and higher ABGA frequency (25% vs 8%; p<0.001) than the comparison group. On prospective analysis, ASO titres were persistently elevated in 57% of patients with Tourette syndrome; however, new infections or newly identified ABGA did not predict clinical exacerbations (all p>0.05). INTERPRETATION Patients with Tourette syndromemight be more prone to GAS infections and develop stronger antibody responses to GAS, probably as a result of underlying immune dysregulation. New GAS infections are unlikely to exert, years after their onset, a major effect upon the severity of neuropsychiatric symptoms.

Tourette syndrome (TS) is a childhood-onset tic disorder associated with abnormal development of brain networks involved in the sensory and motor processing. An involvement of immune mechanisms in its pathophysiology has been proposed. Animal models based on active immunization with bacterial or viral mimics, direct injection of cytokines or patients' serum anti-neuronal antibodies, and transgenic approaches replicated stereotyped behaviors observed in human TS. A crucial role of microglia in the neural-immune crosstalk within TS and related disorders has been proposed by animal models and confirmed by recent post mortem studies. With analogy to autism, genetic and early life environmental factors could foster the involvement of immune mechanisms to the abnormal developmental trajectories postulated in TS, as well as lead to systemic immune dysregulation in this condition. Clinical studies demonstrate an association between TS and immune responses to pathogens like group A Streptococcus (GAS), although their role as risk-modifiers is still undefined. Overactivity of immune responses at a systemic level is suggested by clinical studies exploring cytokine and immunoglobulin levels, immune cell subpopulations, and gene expression profiling of peripheral lymphocytes. The involvement of autoantibodies, on the other hand, remains uncertain and warrants more work using live cell-based approaches. Overall, a body of evidence supports the hypothesis that disease mechanisms in TS, like other neurodevelopmental illnesses (e.g. autism), may involve dysfunctional neural-immune cross-talk, ultimately leading to altered maturation of brain pathways controlling different behavioral domains and, possibly, differences in organising immune and stress responses. This article is part of a Special Issue entitled SI: Neuroimmunology in Health And Disease.

Tourette syndrome (TS) is a childhood-onset tic disorder associated with abnormal development of brain networks involved in the sensory and motor processing. An involvement of immune mechanisms in its pathophysiology has been proposed. Animal models based on active immunization with bacterial or viral mimics, direct injection of cytokines or patients' serum anti-neuronal antibodies, and transgenic approaches replicated stereotyped behaviors observed in human TS. A crucial role of microglia in the neural-immune crosstalk within TS and related disorders has been proposed by animal models and confirmed by recent post mortem studies. With analogy to autism, genetic and early life environmental factors could foster the involvement of immune mechanisms to the abnormal developmental trajectories postulated in TS, as well as lead to systemic immune dysregulation in this condition. Clinical studies demonstrate an association between TS and immune responses to pathogens like group A Streptococcus (GAS), although their role as risk-modifiers is still undefined. Overactivity of immune responses at a systemic level is suggested by clinical studies exploring cytokine and immunoglobulin levels, immune cell subpopulations, and gene expression profiling of peripheral lymphocytes. The involvement of autoantibodies, on the other hand, remains uncertain and warrants more work using live cell-based approaches. Overall, a body of evidence supports the hypothesis that disease mechanisms in TS, like other neurodevelopmental illnesses (e.g. autism), may involve dysfunctional neural-immune cross-talk, ultimately leading to altered maturation of brain pathways controlling different behavioral domains and, possibly, differences in organising immune and stress responses. This article is part of a Special Issue entitled SI: Neuroimmunology in Health And Disease.