Mice treated with the psychostimulant methamphetamine (MA) showed the appearance of intracellular inclusions in the nucleus of medium sized striatal neurones and cytoplasm of neurones of the ...substantia nigra pars compacta but not in the frontal cortex. All inclusions contained ubiquitin, the ubiquitin activating enzyme (E1), the ubiquitin protein ligase (E3‐like, parkin), low and high molecular weight heat shock proteins (HSP 40 and HSP 70). Inclusions found in nigral neurones stained for α‐synuclein, a proteic hallmark of Lewy bodies that are frequently observed in Parkinson's disease and other degenerative disorders. However, differing from classic Lewy bodies, MA‐induced neuronal inclusions appeared as multilamellar bodies resembling autophagic granules. Methamphetamine reproduced this effect in cultured PC12 cells, which offered the advantage of a simple cellular model for the study of the molecular determinants of neuronal inclusions. PC12 inclusions, similar to those observed in nigral neurones, were exclusively localized in the cytoplasm and stained for α‐synuclein. Time‐dependent experiments showed that inclusions underwent a progressive fusion of the external membranes and developed an electrodense core. Inhibition of dopamine synthesis by α‐methyl‐p‐tyrosine (αMpT), or administering the antioxidant S‐apomorphine largely attenuated the formation of inclusions in PC12 cells exposed to MA. Inclusions were again observed when αMpT‐treated cells were loaded with l‐DOPA, which restored intracellular dopamine levels.
Overweight and obesity in the developmental age has become a public health problem. For this reason, prevention projects must be developed in advance with the aim to involve not only children, but ...their parents as well. Our objective is to evaluate the accuracy of the mothers' perceptions of adolescent nutritional status.
Cross-sectional study. We selected a statistical sample of 3,076 subjects (1,583 males, 1,493 females), 8-9 y-old school-children of 164 3rd-grade elementary school classes from throughout Tuscany, as well as their mothers. The mothers' information was gathered via self-administered questionnaires, while the children were given an eating behaviour survey under the supervision of qualified personnel. Mothers' education level (self-reported) height and weight were collected; children's height and weight were measured. The former were asked how they perceived their children's body image.
A correlation exists between the mothers' perceptions of the nutritional state of their children via the silhouettes and the BMI classes of the children, which is equal to 80% with a kappa-Cohen for agreement equal to 0.58 (SE = 0.02; P < 0.0001). However, no correlation exists between the mothers' responses to the question "In your opinion, is your child ...?" and the child's actual BMI class (the exact percentage correlation is equal to 75%, with a kappa-Cohen for agreement equal to 0.43 SE = 0.014; P < 0.0001).
Mothers have an accurate perception of the nutritional status of their children, correctly choosing the silhouette that corresponds to the child's BMI profile without variation by gender. We can assume that mothers in our sample have a good concept about healthy nutritional status.
The prevalence of overweight and obesity in children is rapidly increasing in many countries. For that it has been interesting to investigate the eating habits of 8-9 y-old Tuscany children by paying ...attention to their meals frequency per day and their food choices in total and in relation to children's Body Mass Index (BMI) classes. In addition we considered some environment factors that could affect the children eating behaviours, such as mother's BMI and their education level.
A statistical sample of 3076 (1583 males, 1493 females), 8-9 year-old school-children was collected; weight and height were measured using standardized personnel and instruments. BMI classes were calculated using Cole et al.'s cutoff for children and adolescents. In order to evaluate the consumption frequency of individual meals and various foods, a Food Frequency Questionnaire (FFQ) was used, which was completed by the children themselves at school. A self-administered questionnaire revealed the weight and height of parents and their educational levels. Three educational levels were established: high, medium and low.
The results showed that 92.3% of children ate breakfast from 4-7 times a week, the vast majority at home, while only 3% declared consuming breakfast never or almost never The most preferred breakfast consisted of milk and biscuits for all children's BMI classes. 95.9% of children reported having midmorning snack at school; fruit juice and tea are the most frequently consumed liquid foods, and pizza, salami sandwiches and pre-packaged snacks are the most frequently consumed solid foods in all BMI classes. 93.6% ate afternoon snack for the most part at home, even if 12% of children reported consuming it elsewhere; fruit juice and tea with pizza, sandwiches and pre-packaged snacks are still the most highly consumed foods by all children's BMI classes. The consumption frequency of breakfast (P < 0.001), mid-morning (P < 0.05) and afternoon snack (P < 0.05) of 8-9 y-old Tuscany children decrease with increase the children's BMI classes. The same tendency may be noted for the consumption frequency of breakfast in relation to mother's BMI (P < 0.05) and their education level (P < 0.05). This data strengthens the thesis that some home environments can affect the children's eating behaviours.
No substantial differences in food choices at the meals analyzed were determined among normal weight, overweight and obese children. Children of normal weight had a greater tendency to consume meals more regularly. Mother's BMI and their education level can have influence on children's eating behaviours.
Abstract The PC12 cell line is commonly used as a tool to understand the biochemical mechanisms underlying the physiology and degeneration of central dopamine neurons. Despite the broad use of this ...cell line, there are a number of points differing between PC12 cells and dopamine neurons in vivo which are missed out when translating in vitro data into in vivo systems. This led us to compare the PC12 cells with central dopamine neurons, aiming at those features which are predictors of in vivo physiology and degeneration of central dopamine neurons. We carried out this comparison, either in baseline conditions, following releasing or neurotoxic stimuli (i.e. acute or chronic methamphetamine), to end up with therapeutic agents which are suspected to produce neurotoxicity ( l -DOPA). Although the neurotransmitter pattern of PC12 cells is close to dopamine neurons, ultrastructural morphometry demonstrates that, in baseline conditions, PC12 cells possess very low vesicles density, which parallels low catecholamine levels. Again, compartmentalization of secretory elements in PC12 cells is already pronounced in baseline conditions, while it is only slightly affected following catecholamine-releasing stimuli. This low flexibility is caused by the low ability of PC12 cells to compensate for sustained catecholamine release, due both to non-sufficient dopamine synthesis and poor dopamine storage mechanisms. This contrasts markedly with dopamine-containing neurons in vivo lending substance to opposite findings between these compartments concerning the sensitivity to a number of neurotoxins.
Cellular inclusions containing ubiquitin and alpha-synuclein were observed in PC12 cells treated with metamphetamine (MA). To study the possible involvement of beta-arrestin in inclusion formation, ...we treated PC12 cells with MA for different times and analyzed the ubiquitin proteosome pathway (UPP). We found that beta-arrestin is ubiquitinated in the MA-treated PC12 cell line. The involvement of beta-arrestin in UPP was further supported by electron microscopy and by confocal microscopy, which documented the presence of beta-arrestin in these Lewy body-like inclusions. Our experiments reveal an interesting and previously unappreciated connection between beta-arrestin and ubiquitination and suggest that beta-arrestin could be involved in the development of the inclusion bodies.
The present study explores whether effects induced by amphetamine derivatives on striatal GABA cells might be connected with effects on dopamine (DA) metabolism. Methamphetamine (METH) and ...3,4-methylenedioxymethamphetamine (MDMA, "ecstasy") were administered to C57Black mice following a dosage regimen in which various doses of both drugs were injected i.p. at 2-h intervals. Neuronal inclusions produced under these experimental conditions were examined under electron microscopy. Drugs reducing DA availability prevented inclusion formation; conversely we observed that increasing DA synthesis or impairing physiological DA degradation enhanced the number of inclusions. The present study indicates that the presence of extracellular striatal DA is essential for the production of subcellular alterations induced by amphetamine derivatives. This is in line with a recent hypothesis connecting striatal DA release with degeneration of striatal GABA neurons.
This study investigated the mechanisms involved in the protective actions exerted by lansoprazole against experimental gastric injury. Following the intraluminal injection of ethanol–HCl, the ...histomorphometric analysis of rat gastric sections demonstrated a pattern of mucosal lesions associated with a significant increase in the mucosal contents of malondialdehyde and 8-
iso-prostaglandin F
2α (indices of lipid peroxidation), as well as a decrease in the levels of mucosal sulfhydryl compounds, assayed as reduced glutathione (GSH). Pretreatment with lansoprazole 90 μmol/kg, given intraduodenally as single dose or once daily by intragastric route for 8 days, significantly prevented ethanol–HCl-induced gastric damage. The concomitant changes in the mucosal levels of malondialdehyde, 8-
iso-prostaglandin F
2α and GSH elicited by ethanol–HCl were also counteracted by lansoprazole. In separate experiments, performed on animals undergoing 2-h pylorus ligation, lansoprazole did not enhance the concentration of prostaglandin E
2, bicyclo-prostaglandin E
2, or nitric oxide (NO) metabolites into gastric juice. Western blot analysis revealed the expression of both type 1 and 2 cyclooxygenase (COX) isoforms in the gastric mucosa of pylorus-ligated rats. These expression patterns were not significantly modified by single-dose or repeated treatment with lansoprazole. Lansoprazole also exhibited direct antioxidant properties by reducing 8-
iso-prostaglandin F
2α generation in an in vitro system where human native low-density lipoproteins were subjected to oxidation upon exposure to CuSO
4. The present results suggest that the protective effects of lansoprazole can be ascribed to a reduction of gastric oxidative injury, resulting in an increased bioavailability of mucosal sulfhydryl compounds. It is also proposed that lansoprazole does not exert modulator effects on the gastric expression of COX isoforms as well as on the activity of NO pathways.
Various studies demonstrated that the neurotransmitter norepinephrine (NE) plays a relevant role in modulating seizures; in particular, a powerful effect consists in delaying the kindling of limbic ...areas such as the amygdala and hippocampus. Given the rich NE innervation of limbic regions, we selected a sensitive trigger area, the anterior piriform cortex, to test whether previous loss of noradrenergic terminals modifies sporadic seizures in rats. The damage to locus coeruleus terminals was produced by using the selective neurotoxin N‐(‐2‐chloroethyl)‐N‐ethyl‐2‐bromobenzylamine (DSP‐4, 60 mg/kg i.p.). In intact rats, bicuculline (a GABA‐A antagonist, 118 pmol) microinfused into this area produced sporadic seizures, while in rats previously injected with DSP‐4, bicuculline determined long‐lasting self‐sustaining status epilepticus. In intact rats, sporadic seizures were accompanied by a marked increase in norepinephrine release in the contralateral piriform cortex, while in locus coeruleus‐lesioned rats this phenomenon was attenuated. While bicuculline‐induced sporadic seizures were prevented by the focal infusion of amino‐7‐phosphonoheptanoic acid (AP‐7, a selective NMDA antagonist), or 1,2,3,4‐tetrahydro‐6‐nitro‐2,3‐dioxo‐benzofquinoxaline‐7‐sulphonamide (NBQX, a selective non‐NMDA antagonist), status epilepticus obtained in norepinephrine‐lesioned rats was insensitive to AP‐7 but was still inhibited by NBQX. By using fluorescent staining for damaged (Fluoro‐Jade B) and intact (DAPI) neurons, as well as cresyl violet, we found that rats undergoing status epilepticus developed neuronal loss in various limbic regions. This study demonstrates a powerful effect of noradrenergic terminals in regulating the onset of limbic status epilepticus and its sensitivity to specific glutamate antagonists.
— The present study investigated the gastroprotective effects of the proton pump inhibitor pantoprazole on gastric mucosal damage induced by ethanol‐HCI in rats. Omeprazole was used as reference ...drug. The morphometric analysis of gastric histological sections revealed that pantoprazole and omeprazole dose‐dependently prevented the necrotic mucosal injury evoked by ethanol‐HCI (ED50= 14.1 and 21.6 μmol/kg. respectively). These effects were associated with a marked increment of Alcian blue recovery from gastric bound mucus (ED50=18.8 and 29.3 μmol/kg, respectively). In addition, both pantoprazole and omeprazole inhibited gastric acid secretion in pylorus‐ligated rats (ED50= 1.5 and 3.3 μmol/kg, respectively). Further experiments indicated that the protective effects of pantoprazole were not modified by L‐365,260 (a gastrin receptor antagonist), suramin (a drug able to interfere with endogenous growth factors), NG‐nilro‐L‐arginine (an inhibitor of nitric oxide synthase) or systemic ablation of capsaicin‐sensitive sensory nerves, whereas they were partly blocked by indomethacin (an inhibitor of prostaglandin synthesis) and fully prevented by N‐ethylmaleimide (a potent blocker of sulfhydryl compounds). The present data provide histomorphometric evidence that: 1) pantoprazole is endowed with gastroprotective properties and is more active than omeprazole in preventing the necrotic mucosal damage induced by ethanol‐HCl; 2) according to the rank order of ED50values, the protective effects of both drugs appear to depend mainly on the enhancement of the gastric mucosal barrier rather than on the inhibition of acid secretion; 3) an increased production of prostaglandins, as well as an increased availability of sulfhydryl radicals at the level of the gastric mucosa may account for the gastroprotective effects of pantoprazole.
In recent years several clinical and research findings have demonstrated the involvement of the presynaptic protein α-synuclein in a variety of neurodegenerative disorders which are known as ...synucleinopathies. Although the function of this protein in the physiology of the cell remains unknown, it is evident that both genetic alterations or a mere overexpression of the native molecule produces a degeneration of nigral dopamine-containing neurons leading to movement disorders, as demonstrated in inherited Parkinson's disease. In the present study, we investigated whether widely abused drugs such as methamphetamine and methylenedioxymethamphetamine (ecstasy), which are known to damage the nigrostriatal dopamine pathway of mice, increase the expression of α-synuclein within dopamine neurons of the substantia nigra pars compacta. The results of this study demonstrate that nigrostriatal dopamine denervation and occurrence of intracellular inclusions in nigral neurons produced by amphetamine derivatives are related to increased expression of α-synuclein within dopamine neurons of the substantia nigra. This lends substance to the hypothesis that increased amounts of native α-synuclein may be per se a detrimental factor for the dopamine neurons.
