Identification in the rat brain of a set of nuclear proteins interacting with H1° mRNA.

Synthesis of H1° histone, in the developing rat brain, is also regulated at post-transcriptional level. Regulation of RNA metabolism depends on a series of RNA-binding proteins (RBPs); therefore, we searched for H1° mRNA-interacting proteins. With this aim, we used in vitro transcribed, biotinylated H1° RNA as bait to isolate, by a chromatographic approach, proteins which interact with this mRNA, in the nuclei of brain cells. Abundant RBPs, such as heterogeneous nuclear ribonucleoprotein (hnRNP) K and hnRNP A1, and molecular chaperones (heat shock cognate 70, Hsc70) were identified by mass spectrometry. Western blot analysis also revealed the presence of cold shock domain-containing protein 2 (CSD-C2, also known as PIPPin), a brain-enriched RBP previously described in our laboratory. Co-immunoprecipitation assays were performed to investigate the possibility that identified proteins interact with each other and with other nuclear proteins. We found that hnRNP K interacts with both hnRNP A1 and Hsc70 whereas there is no interaction between hnRNP A1 and Hsc70. Moreover, CSD-C2 interacts with hnRNP A1, Y box-binding protein 1 (YB-1), and hnRNP K. We also have indications that CSD-C2 interacts with Hsc70. Overall, we have contributed to the molecular characterization of a ribonucleoprotein particle possibly controlling H1° histone expression in the brain.

The effects of a 3-week training on basal biomarkers in professional soccer players during the preseason preparation period.

AIM: The present study aimed at investigating the effect of a 3-week training on biomarkers in professional soccer players during the preseason preparation-period. METHODS: Eight participants (age 22.5±2.2 yrs) were enrolled in the study. During the physical preparation period players have attended a training program (51.9 hours) formulated by coaches of "Equipe-Sicilia-2009". RESULTS: At rest, the lipid profile, the creatine kinase (CK), the lactic-acid dehydrogenase (LDH) and the expression of nuclear receptors peroxisome-proliferator-activated receptors (PPAR α/γ) were analyzed before starting and after 3 weeks of training. The plasma level of CK in our samples showed great variability already in the baseline: value was on average nearly 500 IU/l showed that a large amount of these athletes were a high responders. This biomarker showed a reduction (P<0.01) after 3 weeks of training. No modifications were found in the LDH plasma level, in the lipid profile and in the expression of mRNA of PPAR α/γ and also no significant person's correlations were found among variables. CONCLUSION: In conclusion, we retain that those basal biomarkers, except CK, are not able to assist coaches to better understand training adaptations and overreaching mechanisms during a 3-week of preseason preparation-period. More studies are necessary to confirm these results.

Symbolic function explored in children with epilepsy and headache.

AIM: When the body gets ill, the attack on the ability to think is one of the possible effects that can be observed. The aim of this study is to explore symbolic functions in children with epilepsy and primary headache. METHODS: Fifty five little patients have been recruited from the Neuropsychiatry infant Clinic of the University of Palermo; 48 males (61%) and 27 females (39%), all suffering from epilepsy (53-71%) and primary headache (22-29%), their ages varying from 7 to 11 (average: 9.4+/-1.2). Subjects that had never suffered from chronic or neurological diseases were also recruited as a control group. The control group was levelled with the group to be tested in age and number (75 subjects), age (range: 7-11; average: 9.1+/-9) sex (males: 48-61%; females: 27-39%). The instruments that have been used were: colored progressive matrices in order to measure the development of the cognitive functions; semi-structured interview on dreams in order to estimate the quality of the mentalization; drawing stories technique in order to estimate the quality of the psychological suffering; fairytales method of Düss in order to observe the defensive strategies used by the subjects. RESULTS: The cognitive performances were the same in the two groups. The answers to the semi-structured interview on dreams were different as far as coherent theory on dreams and memory of the last dream and its quality are concerned. The Drawing stories technique and the Fairytales method of Düss with the children in the clinical group - especially the epileptic patients - showed a high presence of psychical suffering, unprocessed or impossible to process. CONCLUSIONS: The results allow to estimate a psychological suffering, focusing onbody sickness and to discriminate some specific ways of constriction of the imaginary, linked to either repression or to splitting/dissociation.

Expressions of discomfort in adolescents. Clinical reflections resulting from an exploratory study of a group of students.

AIM: The aim of this study was to explore some fundamental dimensions of adolescent discomfort relating to food, the body, sexuality, relationships in general and health care. METHODS: We recruited 206 students (66 males, 140 females; average age=19.4+/-2) and we administered 4 questionnaires: the first two to examine their relationship with food, the third to explore adolescents' attitudes to their social context, sexuality and first sexual experiences; the fourth to measure their ability to look after their health. RESULTS: The results show the different role that sexual styles have in the expression of juvenile discomfort. Girls tend to live the discomfort by mentalizing it in relationships and sexuality and contextualizing it in their bodies; boys direct their discomfort towards occasional drug taking or drug addiction, or alcohol abuse, the excitement of speed, or dangerous driving. CONCLUSIONS: In clinical work with adolescents it seems appropriate to refer to the relational gradient of risk behaviours, that is their linking function within their peer group. Behaviours with a high relational gradient, which develop within the group or are related to the group, in most cases are normal modalities for facing the important challenges of adolescence. These behaviours are symptoms of the normal process of adolescent development. We should pay particular attention to behaviours with a low relational gradient because they indicate a possible interruption in their development.

Predictive signs and indicators of aggressiveness and violence: a comparison between a group of adolescents attending an external penal area, a group of prisoners and a group of patients with borderline personality disorder.

AIM: Aggressive and violent behaviours in juveniles and young adults have become increasingly widespread. Most such behaviours have 2 common roots: lack of actual motivation and brutality. The most reliable indicators and predictive signs have been linked to structural personality features (e.g. defensive strategies, impulse control). With this cross-sectional study we wanted to determine specific indicators and predictive signs of violent and aggressive behaviours in these population segments. METHODS: We compared the structural personality features of 3 groups: one with 26 male adolescents with conduct disorder (F91.8), one with 29 male patients with borderline personality disorder (F60.31) and one with 34 male prisoners with antisocial personality disorder (F60.2). The test battery included: the ''Structured clinical interview for DSM-IV axis II disorders'' (for the recruitment of adult groups); the ''Indicators of aggressive conduct'' and the ''Profile of mood states'' (for the recruitment of the adolescent group); the ''Defense mechanisms inventory'' (DMI). RESULTS: The predominant defensive strategies and starting emotional backgrounds differed significantly among the groups. A profile of low emotional response was found to support aggressiveness, while a profile of aggressiveness and violence was associated with clinically significant impulsivity. CONCLUSIONS: Several indicators and clinical predictors of aggressiveness and violence, as well as high-risk-profiles, may be determined using these instruments.

Functional feature of a novel model of blood brain barrier: studies on permeation of test compounds.

Drug delivery to the central nervous system (CNS) is subject to the permeability limitations imposed by the blood-brain barrier (BBB). Several systems in vitro have been described to reproduce the physical and biochemical behavior of intact BBB, most of which lack the feature of the in vivo barrier. We developed a fully formed monolayer of RBE4.B immortalized rat brain microvessel endothelial cells (ECs), grown on top of polycarbonate filter inserts with cortical neuronal cells grown on the outside. Neurons induce ECs to synthesize and sort occludin to the cell periphery. Occludin localization is regulated by both compositions of the substratum and soluble signals released by cortical co-cultured neurons. The observed effects do not require strict physical contact among cells and neurons. To assess the physiological function of the barrier we examined the transendothelial transfer of three test compounds: dopamine, L-tryptophan and L-DOPA. Polycarbonate filter inserts, where ECs were co-cultured with neurons, were assumed as open two compartments vertical dynamic models. Permeation studies demonstrated that the ECs/neurons co-cultures possess permeability characteristics approaching those of a functional BBB: the system behaved as a selective interface that excludes dopamine permeation, yet permits L-tryptophan and L-DOPA to cross. The movement of test compounds from the donor to the acceptor compartment was observed at a distinct time from the start of co-culture. Transfer was determined using standard kinetic equations. Different performance was observed after 5 and 7 days of co-culture. After 5 days dopamine, L-tryptophan and L-DOPA passively permeate through the membrane as indicated by fittings with a first-order kinetic process equation. After 7 days of co-culture, occludin localizes at ECs periphery, dopamine does not cross the barrier to any further extent, while the transfer of L-tryptophan and L-DOPA fits well with a saturable Michaelis-Menten kinetic process, thus indicating the involvement of a specific carrier-mediated transport mechanism. Permeation studies confirmed that culture of ECs in the presence of neurons induces the characteristic permeability limitations of a functional BBB.

Neurons and ECM regulate occludin localization in brain endothelial cells.

We report that extracellular matrix and neurons modulate the expression of occludin, one of the main components of tight junctions, by rat brain endothelial cells (RBE4.B). Of the three extracellular matrix proteins which we tested (collagen I, collagen IV, and laminin), collagen IV stimulated at the best the expression of occludin mRNA. The corresponding protein, however, was not synthesized. Significant amounts of occludin accumulated only when RBE4.B cells were cultured on collagen IV-coated inserts, in the presence of cortical neurons, plated on laminin-coated companion wells. Finally, occludin segregated at the cell periphery, only when endothelial cells were co-cultured with neurons for > or = 1 week.