Healthy lifestyle promotion in primary schools through the board game Kaledo: a pilot cluster randomized trial.

The board game Kaledo was proven to be effective in improving nutrition knowledge and in modifying dietary behavior in students attending middle and high school. The present pilot study aims to reproduce these results in younger students (7-11 years old) attending primary school. A total of 1313 children from ten schools were recruited to participate in the present study. Participants were randomized into two groups: (1) the treatment group which consisted of playing Kaledo over 20 sessions and (2) the no intervention group. Anthropometric measures were carried out for both groups at baseline (prior to any treatment) and at two follow-up post-assessments (8 and 18 months). All the participants completed a questionnaire concerning physical activity and a 1-week food diary at each assessment. The primary outcomes were (i) BMI z-score, (ii) scores on physical activity, and (iii) scores on a dietary questionnaire. BMI z-score was significantly lower in the treated group compared to the control group at 8 months. Frequency and duration of self-reported physical activity were also significantly augmented in the treated group compared to the control group at both post-assessments. Moreover, a significant increase in the consumption of healthy food and a significant decrease in junk food intake were observed in the treated group. CONCLUSION: The present results confirm the efficacy of Kaledo in younger students in primary schools, and it can be used as a useful nutritional tool for obesity prevention programs in children. What is Known: • Kaledo is a new educational board game to improve nutrition knowledge and to promote a healthy lifestyle. • In two cluster randomized trials conducted in Campania region (Italy), we showed that Kaledo could improve nutrition knowledge and dietary behavior and have a positive effect on the BMI z-score in children with age ranging from 9 to 14 years old attending school. • Kaledo may be used as an effective tool for obesity prevention programs in middle and high school students. What is New: • Investigating the effects of Kaledo on younger primary school children (7-11 year olds), Kaledo could be an effective tool in obesity prevention programs for children as young as 7 years old.

Epigenetics and cortical spreading depression: changes of DNA methylation level at retrotransposon sequences.

Cortical spreading depression (CSD) is an evolutionarily conserved phenomenon that involves a slow and self-propagating depolarization wave associated with spontaneous depression of electrical neuronal activity. CSD plays a central role in the pathophysiology of several brain diseases and is considered to be able to promote "Preconditioning". This phenomenon consists of the brain protecting itself against future injury by adaptation. Understanding of the molecular mechanisms underlying Preconditioning has significant clinical implications. We have already proposed that the long-lasting effects of CSD could be related to silencing of retrotransposon sequences by histone methylation. We analyzed DNA methylation of two retrotransposon sequences, LINE1 and L1, and their corresponding expression pattern after CSD induction. Based on immunoprecipitation assay of the methylated DNA (meDIP), we demonstrated hypermethylation of both sequences in preconditioned rat brain cortex compared with a control 24 h after CSD induction. Using quantitative PCR, we also showed that CSD induction caused a decrease of the transcript level of both retrotransposon sequences. Our data are consistent with the hypothesis of epigenetic modifications in Preconditioning-dependent neuroprotection by increasing genome stability via the silencing of retrotransposon sequences.

Kaledo, a board game for nutrition education of children and adolescents at school: cluster randomized controlled trial of healthy lifestyle promotion.

UNLABELLED: During childhood and adolescence, a game could be an effective educational tool to learn healthy eating habits. We developed Kaledo, a new board game, to promote nutrition education and to improve dietary behavior. A two-group design with one pre-treatment assessment and two post-treatment assessments was employed. A total of 3,110 subjects (9-19 years old) from 20 schools in Campania, Italy, were included in the trial. In the treated group, the game was introduced each week over 20 consecutive weeks. Control group did not receive any intervention. The primary outcomes were (i) score on the "Adolescent Food Habits Checklist" (AFHC), (ii) scores on a dietary questionnaire, and (iii) BMI z-score. At the first post-assessment (6 months), the treated group obtained significantly higher scores than the control group on the AFHC (14.4 (95 % confidence interval (CI) 14.0 to 14.8) vs 10.9 (95 % CI 10.6 to -11.2); F(1,20) = 72.677; p < 0.001) and on four sections of the dietary questionnaire: "nutrition knowledge" (6.5 (6.4 to 6.6) vs 4.6 (4.5 to 4.7); F(1,16) = 78.763; p < 0.001), "healthy and unhealthy diet and food" (11.2 (11.0 to 11.4) vs 10.4 (10.3 to 10.6); F(1,32) = 21.324; p < 0.001), "food habits" (32.4 (32.0 to 32.8) vs 27.64 (27.3 to 28.0); F(1,26) = 195.039; p < 0.001), and "physical activity" (13.4 (13.2 to 13.7) vs 12.0 (11.8 to 12.6); F(1,20) = 20.765; p < 0.001). Moreover, the treated group had significantly lower BMI z-score with respect to the controls at the first (0.44 (0.42 to 0.46) vs 0.58 (0.56 to 0.59), F(1,18) = 16.584, p = 0.001) and at the second (18 months) (0.34 (0.30 to 0.38) vs 0.58 (0.54 to 0.62), F(1,13) = 7.577; p = 0.017) post-assessments. CONCLUSION: Kaledo improved nutrition knowledge and dietary behavior over 6 months and had a sustained effect on the BMI z-score. Therefore, it may be used as an effective tool in childhood and adolescence obesity prevention programs.

Cortical spreading depression increases the phosphorylation of AMP-activated protein kinase in the cerebral cortex.

Cortical spreading depression (CSD) enhances ischemic tolerance to temporary focal ischemia. Although this effect most likely requires the expression or activation of neuroprotective factors, their identity remains relatively unknown. One important factor involved in neuroprotection is adenosine monophosphate-dependent kinase (AMPK), a serine/threonine kinase that is activated via phosphorylation. This activation occurs in response to brain ischemia, hypoxia, or glucose deprivation. Thus, to determine the potential mechanism of the neuroprotective effects of CSD, we tested whether AMPK becomes phosphorylated into phospho-AMP-activated protein kinase (pAMPK) after CSD. CSD was induced for 15 min in three groups of five rats. The animals were subsequently sacrificed after 2, 4 or 24 h. Western blot analyses were performed to determine the AMPKα and pAMPKα levels in the cortex (right and left hemispheres), and immunohistochemistry and immunofluorescence were performed to determine the localisation of AMPKα and pAMPKα in the cerebral cortex. These results demonstrated a significant increase in pAMPKα at 24 h (but not at 2 and 4 h) after CSD. In contrast, un-phosphorylated AMPK expression did not change. The increase in pAMPKα was confined to neurons (predominantly neurons located in the superficial layers of the cerebral cortex) and was not observed in astroglial cells. Taken together, these data indicate that AMPK is activated by CSD, and suggest that this activation may contribute to the neuroprotective effect of CSD.

Retrospectively and prospectively modulated hippocampal place responses are differentially distributed along a common path in a continuous T-maze.

Hippocampal place responses can be prospectively or retrospectively modulated by the animal's future or prior trajectory. Two main hypotheses explain this. The "multiple-map hypothesis" switches between different maps for different trajectories (rate remapping). In contrast, in the "buffer hypothesis," the hippocampus encodes an ongoing representation that includes the recent past and/or the impending future choice. This study examines the distribution of prospective and retrospective responses distributed along a common path in a continuous T-maze (providing all four combinations of provenance and destination) during a visual discrimination task. The multiple-map hypothesis predicts either uniform distributions or concerted shifts about a task-decision relevant point, whereas the buffer hypothesis predicts a time-limited overexpression around choice points (with retrospective responses after the central arm entry point and prospective responses nearer its exit). Here bilateral recordings in the dorsal CA1 region of the rat hippocampus show that retrospective responses were twice as prevalent as prospective responses. Furthermore, retrospective and prospective modulations have distinct spatial distributions, with retrospective primarily in the first two-thirds of the central arm and prospective restricted to the last third. To test for possible trial-by-trial remapping in relation to the two-thirds transition point, data from the first and second halves of the sessions were compared. Backward drift of path-modulated activity was significant only for retrospective, but not prospective, fields. Thus, these data are more consistent with the buffer hypothesis. Retrospective and prospective modulation would then participate in a single hippocampal representation of spatial and behavioral context.

Methionine-enriched diet decreases hippocampal antioxidant defences and impairs spontaneous behaviour and long-term potentiation in rats.

Diets high in methionine lead to elevation of plasma homocysteine levels which are possibly linked to neurodegenerative diseases and oxidative stress. In the present study, we investigated the effects of methionine-enriched diet on antioxidant defences, on rat spontaneous behaviour and on the ability to sustain long-term potentiation in the dentate gyrus (DG). Sprague-Dawley rats were fed either a standard laboratory diet or a methionine enriched-diet (1% or 5% methionine in drinking water) for 8 weeks. After the 8 weeks, the animals were tested for spontaneous motor activity and habituation in an open field maze, for anxiety-like behaviour in an elevated plus maze and for the ability to sustain long-term potentiation (LTP) induced in the dentate gyrus under urethane anaesthesia. The brains were then removed and histochemically stained for superoxide dismutase (SOD) activity. Rats fed on 5% methionine significantly reduced total distance travelled during the open field test and exhibited no habituation with respect to the other two groups. Rats fed on 5% methionine also showed a significant increase of the anxiety level. Moreover, in this group, the ability to induce LTP in DG was impaired. SOD activity was significantly increased in the cerebral cortex of the rats fed on 1% and 5% methionine with respect to the control group. In conclusion, 5% methionine in drinking water led to evident impairment of locomotor skills and of synaptic plasticity. SOD activity in the cortex was increased in both the groups fed on 1% and 5% methionine, thus suggesting that metabolic adjustments, triggered by the methionine-enriched diet, are likely mediated by reactive oxygen species.

Dynamics of decision-related activity in hippocampus.

Place-selective activity in hippocampal neurons can be modulated by the trajectory that will be taken in the immediate future ("prospective coding"), information that could be useful in neural processes elaborating choices in route planning. To determine if and how hippocampal prospective neurons participate in decision making, we measured the time course of the evolution of prospective activity by recording place responses in rats performing a T-maze alternation task. After five or seven alternation trials, the routine was unpredictably interrupted by a photodetector-triggered visual cue as the rat crossed the middle of central arm, signaling it to suddenly change its intended choice. Comparison of the delays between light cue presentation and the onset of prospective activity for neurons with firing fields at various locations after the trigger point revealed a 420 ms processing delay. This surprisingly long delay indicates that prospective activity in the hippocampus appears much too late to generate planning or decision signals. This provides yet another example of a prominent brain activity that is unlikely to play a functional role in the cognitive function that it appears to represent (planning future trajectories). Nonetheless, the hippocampus may provide other contextual information to areas active at the earliest stages of selecting future paths, which would then return signals that help establish hippocampal prospective activity.

Reconstruction of field excitatory post-synaptic potentials in the dentate gyrus from amperometric biosensor signals.

A new feasible and reproducible method to reconstruct local field potentials from amperometric biosensor signals is presented. It is based on the least-square fit of the current response of the biosensor electrode to a voltage step by the use of two time constants. After determination of the electrode impedance, Fast Fourier Transform (FFT) and Inverse FFT are performed to convert the recorded amperometric signals into voltage and trace the local field potentials using a resistor-capacitor circuit-based model. We applied this method to reconstruct field evoked potentials from currents recorded by a lactate biosensor in the rat dentate gyrus after stimulation of the perforant pathway in vivo. Initial slope of the reconstructed field excitatory postsynaptic potentials was used in order to demonstrate long term potentiation induced by high frequency stimulation of the perforant path. Our results show that reconstructing evoked potentials from amperometric recordings is a reliable method to obtain in vivo electrophysiological and amperometric information simultaneously from the same electrode in order to understand how chemical compounds vary with and modulate the dynamics of brain activity.

Plasma glutamine decreases immediately after surgery and is related to incisiveness.

Glutamine (gln) is the most abundant free amino acid in the blood. It is involved in important metabolic and biochemical processes, like cell proliferation and oxidative stress. Previous studies have demonstrated that gln concentration in human plasma decreases in several conditions such as sepsis, ischemia-reperfusion, trauma, major surgery and burn. The aim of the present work was to compare the acute effects of different types of surgical interventions and of anesthetization on blood gln concentration. Plasma samples from 88 subjects (30 males and 58 females) were collected before and after major or minor surgery and the gln concentration was analyzed with high-performance liquid chromatography. The results showed that plasma gln concentration after surgery was lower than pre-surgery values and that in major surgery the decrease of gln was higher than in minor surgery. No significant effect was shown for sex or type of anesthesia. These results demonstrate the importance of a gln supplementation before a surgical intervention and show that the amount of gln supplementation should also be adjusted based on the type of surgery.

Cortical spreading depression affects reactive oxygen species production.

Cortical spreading depression (CSD) is a reversible electrophysiological phenomenon that is not associated with tissue damage under normal blood supply. The induction of CSD during ischemia causes an increase in tissue damage, while pre-conditioning with CSD decreases the damage induced by a subsequent episode of ischemia. The mechanisms underlying these effects are not clear. Because the production of reactive oxygen species (ROS) is involved in tissue damage during ischemia-reperfusion, the aim of the present study was to evaluate the effects of CSD on superoxide production (O(2)(-)), on hydrogen peroxide (H(2)O(2)) production and on superoxide dismutase (SOD) activity in the cerebral cortex. CSD was induced by KCl application on the cortical surface in rats. O(2)(-) production was evaluated using dihydroethidium (DHE) that is oxidized to the fluorescent dye ethidium (HEt) by O(2)(-). The extracellular level of H(2)O(2) was evaluated by microdialysis sampling and HPLC analysis. SOD activity was evaluated with a histochemical assay. The results showed an increase in H(2)O(2) production, an increase in SOD activity and a decrease in O(2)(-) concentration 1h after CSD induction.

Persistent facial pain increases superoxide anion production in the spinal trigeminal nucleus.

Previous studies have demonstrated that there is an increase in oxidative stress in the cerebral cortex of rats after repeated painful stimulation and that long-lasting pain increases the production of superoxide ion (O(2) (-)), nitric oxide and peroxynitrite due to the activation of AMPA and NMDA receptors. The purpose of the present study was to evaluate the possible role of O(2) (-) in the transmission of oro-facial pain. Formaldehyde 1% was injected subcutaneously into one vibrissal pad of adult male Sprague-Dawley rats as a model of persistent pain, then O(2) (-) production and superoxide dismutase (SOD) activity were evaluated in the left and right spinal trigeminal nuclei. O(2) (-) production was revealed using dihidroetidium (DHE) injected at 10 or 45 min after the formalin injection in conscious or anaesthetized rats. A histochemical assay for SOD was performed to evaluate the activity of SOD at 10 min after the formalin injection. The results showed a significant increase in O(2) (-) production in the homolateral nucleus at 45 min. However, there was no significant difference between the two sides at 10 min after the formalin injection. No significant difference was observed in SOD activity between the two sides of the spinal trigeminal nucleus. This study demonstrated that there is an increased production of O(2) (-) in the second phase but not in the first phase of the formalin test; thus O(2) (-) is involved in pain induced by inflammation, but not in acute pain.

Fast and low-cost analysis of heart rate variability reveals vegetative alterations in noncomplicated diabetic patients.

The aim of the present preliminary study was to describe a simple protocol for the analysis of the heart rate variability (HRV) that can reveal the different autonomic control between noncomplicated diabetic patients and normal subjects within 15 min. The power spectrum of the HRV was evaluated on 5-min-long electrocardiographic recordings in both the supine and the seated positions in 30 noncomplicated non-insulin-dependent diabetic (NIDDM) patients and in 30 healthy volunteers. In healthy subjects the low-frequency (LF) value was higher in seated position than in supine position, while in diabetic patients the LF value in seated position did not differ from that in supine position and did not differ from that in healthy subjects in supine position. The present work demonstrates that the protocol described reveals a different autonomic regulation of the heart rate in healthy subjects and in NIDDM patients even if there is no clinically evident autonomic neuropathy.

Peripheral amino Acid levels in schizophrenia and antipsychotic treatment.

Abnormal levels of amino acids have been reported in patients with schizophrenia and have also been investigated as a biomarker to monitor antipsychotic treatment, however results have been inconsistent. The purpose of the present review is to summarize the evidence in the literature of whether amino acid levels can be a biomarker and predict the treatment outcome in schizophrenia. The current review does not support amino acid concentration as a useful biomarker for monitoring antipsychotic response in patients with schizophrenia, although there is evidence that high levels of serum homocysteine and glutamate might be considered as a trait marker for schizophrenia. This review has also highlighted a considerable dearth of studies, specifically of studies evaluating antipsychotic side-effects.

Kalèdo, a new educational board-game, gives nutritional rudiments and encourages healthy eating in children: a pilot cluster randomized trial.

INTRODUCTION: Prevention of obesity and overweight is an important target for health promotion. Early prevention requires an intervention during childhood and adolescence. At these stages, the game could be an appropriate means to teach nutrition knowledge and to influence dietary behaviour. To this end, the authors developed Kalèdo, a new board-game. OBJECTIVE: The aim of the present study was to test the efficacy of Kalèdo on changes in nutrition knowledge and dietary behaviour in a pilot study conducted in three middle schools in Naples, Italy. MATERIALS AND METHODS: A simple two-group design (treatment and control) with pre- and post-assessment was employed. The classroom was the unit of recruitment and random assignment to groups. All students (307) in the participating schools were invited to participate. Data analysis was performed on 241 subjects. During 24 weeks, a group of 153 children from 8 classrooms (11-14 year old Caucasian subjects; 78 male, 75 female) was involved in 15-30 minute-long play sessions once a week. A questionnaire was given to the participants at the beginning and at the end of the study to evaluate nutrition knowledge (31 questions), physical activity (8 questions) and food intake (34 questions). Anthropometric measurements were also carried out. A second group of 88 children from 5 classrooms (same age and ethnicity; 55 male, 33 female) was investigated at the same times with the same questionnaire and anthropometric measures but they did not receive any play sessions with Kalèdo. OBSERVATION: Children playing Kalèdo showed a significant increase in nutrition knowledge (p<0.05) and in weekly vegetable intake (p<0.01) with respect to the control. CONCLUSION: The results suggest that Kalèdo could be an effective instrument to teach children about healthy diet. More research is needed to study the long term effect of this intervention.

Annurca apple-rich diet restores long-term potentiation and induces behavioral modifications in aged rats.

The impairment of brain functions during aging could be prevented by an increase in antioxidative defenses. In fact, dietary manipulation with antioxidants or with antioxidant-rich extracts from vegetables and fruit can ameliorate age-associated behavioral alterations. Moreover, dietary supplements of antioxidants can reverse the age-dependent impairment to sustain long-term potentiation (LTP). There is evidence that fresh fruit could be more effective than antioxidant supplements or fruit and vegetable extracts but the effect of a diet enriched in fresh fruit on brain aging process has not been investigated until now. The aim of the present study was to verify whether dietary manipulation with fresh apples could influence age-induced changes in motor and emotional behavior and in LTP in rats. Groups of aged and young rats were fed with a standard diet or a diet supplemented with fresh apples of annurca variety for 10 weeks and assessed for behavioral performance in the open field and elevated plus maze tests and for the ability to sustain LTP. The brains were then removed, histochemically stained for superoxide dismutase (SOD) and SOD activity was measured by computer-assisted image analysis. The aged rats fed with the enriched diet showed a significant decrease in the anxiety level. Moreover, they improved in the ability to sustain LTP, reaching the level of the young rats. SOD activity was increased in the aged rats fed with the standard diet whereas SOD activity in the hippocampus of the aged rats treated with annurca apple was at the level of the young animals. These results suggest that a diet rich in annurca apple could have an important role in health-care during aging.

Trigeminal pain transmission requires reactive oxygen species production.

Three experiments were conducted in order to investigate the possible involvement of the reactive oxygen species in the nociception within the subnucleus caudalis of the spinal trigeminal nucleus (Vc). In the first experiment the extracellular level of hydrogen peroxide was evaluated by microdialysis in the Vc of two groups of six rats before and after a formalin (group 1) or saline solution (group 2) injection into the upper lip. In the second experiment the formalin test was conducted in three groups of 6 rats after a microinjection of 2-methoxyestradiol (2-ME, a superoxide-dismutase inhibitor; group 1) or N-acetylcysteine (NAC, an oxygen intermediate scavenger; group 2) or saline solution (group 3) into the Vc. In the third experiment an histochemical assay for superoxide dismutase activity was performed on two groups of 4 rats each 2 h after a formalin (group 1) or saline solution (group 2) injection into the upper lip. The results showed that (1) the level of hydrogen peroxide increases into the Vc during facial pain (134% of baseline); (2) the inhibition of superoxide dismutase or the removal of oxygen intermediate within the Vc decreases the sensibility to facial pain stimuli; and (3) persistent facial pain stimuli decrease the superoxide activity into the Vc (90% of counter-lateral). These data indicate that reactive oxygen species are produced in the Vc during persistent facial pain and are necessary for the transmission of pain.

Evidence that GABAergic neurons in the spinal trigeminal nucleus are involved in the transmission of inflammatory pain in the rat: a microdialysis and pharmacological study.

The aim of this experiment was to investigate the role of the gamma-aminobutyric acid (GABA)-ergic transmission in the nociception within the spinal trigeminal nucleus. The formalin test was used as an animal model of inflammatory pain. Two groups of six rats were used. The behavioural response to the labial injection of formaldehyde (50 microl of a 5% solution) (group 1) or saline (group 2) was evaluated by recording the time spent in facial grooming during a period of 8 min (one period before and seven consecutive periods after the injection). The extracellular concentration of GABA in the trigeminal caudalis nucleus was evaluated, during the formalin test, on samples of 30 microl each (one sample before and three samples after the labial injection) obtained by microdialysis and analysed by HPLC with electrochemical detection of the o-phtalaldeyde pre-column derivate. Subsequently, three more groups of six rats each were injected with saline, muscimol (GABAa receptor agonist), or bicuculline (GABAa receptor antagonist) in the trigeminal caudalis nucleus, before performing the formalin test. The injection of formaldehyde induced a biphasic behavioural response and an increase of the GABA levels at 15-45 min. The injection of bicuculline, but not muscimol or saline, strongly decreased the behavioural response of the formalin test. These findings suggest that GABAergic neurons in the trigeminal caudalis nucleus are involved in the transmission of nociceptive information.

Quantitative histochemical assay for superoxide dismutase in rat brain.

Superoxide anions are highly reactive radicals overproduced in many pathological situations such as inflammation and ischemia. One of the major factors in the protection from superoxide anions is the enzyme superoxide dismutase (SOD), which catalyzes the dismutation of superoxide to hydrogen peroxide. This study presents a quantitative histochemical method to estimate SOD activity in rat brain tissue sections. This method is based on the cerium capture method and 3,3'-diaminobenzidine amplification of transition cerium compounds. Substrate for SOD was provided by reduction of oxygen during the autoxidation of riboflavin in the presence of UV light. This histochemical method reveals the overall activity of the three different forms of SOD described in mammalian tissues: cytosolic copper-zinc SOD, mitochondrial manganese SOD, and the high molecular weight extracellular SOD. Eventually, this method can be used to quantify SOD activity in tissue sections by image analysis.