Monitoraggio real-time della neurochimica corticale in topi liberi di muoversi durante il sonno, la privazione di sonno e la veglia medinte microdialisi in vivo e neurosensori amperometrici

Aim: for millennia, mankind has tried to understand the function and the importance of sleep and this is a mystery that biology has yet to solve. The purpose of this research was study neurochemistry during sleep, sleep deprivation and wakefulness by means of in-vivo microdialysis and amperometric neurosensors. Method: the neurochemistry of the brain was studied by the combination of electrochemical and microdialysis techniques, electroencephalographic (EEG) and electromyographic (EMG) recordings. The mice were implanted with three electrodes for electroencephalographic recordings and two microdialysis probes. During the experiment, samples were taken every 15 minutes and then analyzed by UHPLC/CD ORBITRAP that allows to analyze all the molecules present the sample. Moreover, the biochemical dynamics of Ascorbic Acid (AA) have been extensively studied both in vitro and in vivo by the development of a nanostructured amperometric microsensor decorated with multiwall carbonanotubes (MWCNTs). Results: through multiple experiments it was possible to study the variation in levels of cortical AA in freely moving mice during sleep and sleep deprivation, furthermore, the microdialysis technique, combined with EEG and EMG, allowed to monitor extracellular molecules (as neurotransmitters) that are involved in the neurochemistry of sleep, sleep deprivation and wakefulness. Conclusion: understand the biology that is the basis of sleep is the goal of many researchers. It seems to be involved in different processes such as learning and memory, also, his disorders are the basis of diseases (i.e. depression). This study allows, for the first time, to have an overview of the neurochemistry of sleep contribute, therefore, to better understanding of its mechanisms and functions and laying the groundwork for future studies

Editorial: Emerging heterocycles as bioactive compounds

Heterocycles represent a privileged scaffold due to their ability to interact with biological systems via heteroatoms. It is no coincidence that every year the Food and Drug Administration approves numerous new drugs that contain at least one heterocyclic system as active pharmacophoric part in their structure. Many heterocyclic compounds with therapeutic properties, including anticancer, antimicrobial, anti-inflammatory and so on, come from natural sources such as plants and animals, and medicinal chemists very often use them to study their chemical space and improve their biological activity. In fact, several efficient approaches for the formation of aromatic heterocyclic compounds and their derivatives have been described in the literature in the past, but the development of new green synthetic procedures and methodologies for their high-yield synthesis is increasingly in demand in drug discovery program. This Editorial collects recent research progress in the field of medicinal chemistry focused on the synthesis of new bioactive heterocycles of different types of activities, including anticancer, antibacterial and anti-inflammatory

Productivity changes in the Mediterranean Sea drive foraging movements of yelkouan shearwater Puffinus yelkouan from the core of its global breeding range

Pelagic seabirds are tied to their breeding colonies throughout their long-lasting breeding season, but at the same time, they have to feed in a highly dynamic marine environment where prey abundance and availability rapidly change across space and seasons. Here, we describe the foraging movements of yelkouan shearwater Puffinus yelkouan, a seabird endemic to the Mediterranean Sea that spends its entire life cycle within this enclosed basin and whose future conservation is intimately linked to human-driven and climatic changes affecting the sea. The aim was to understand the main factors underlying the choice of foraging locations during the reproductive phases. A total of 34 foraging trips were obtained from 21 breeding adults tagged and tracked on Tavolara Archipelago (N Sardinia, Italy). This is the largest and most important breeding area for the species, accounting for more than 50% of the world population. The relationships between foraging movements during two different breeding stages and the seasonal changes of primary productivity at sea were modeled. Movements appeared to be addressed toward inshore (<20 km), highly productive, and relatively shallow (<200 m) foraging areas, often in front of river mouths and at great distances from the colony. During incubation, the Bonifacio Strait and other coastal areas close to North and West Sardinia were the most preferred locations (up to 247 km from the colony). During the chick-rearing phase, some individuals reached areas placed at greater distances from the colony (up to 579 km), aiming at food-rich hotspots placed as far north as the Gulf of Lion (France). The need for such long distance and long-lasting foraging trips is hypothesized to be related to unfavorable conditions on the less productive (and already depleted) Sardinian waters

The neurotoxin 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine induces apoptosis in mouse nigrostriatal glia. Relevance to nigral neuronal death and striatal neurochemical changes.

Swiss mice were given 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP), 25 mg/kg/day, for 5 consecutive days and killed at different days after MPTP discontinuance. Decreases in striatal tyrosine hydroxylase activity and levels of dopamine and its metabolites were observed 1 day after MPTP discontinuance. Ascorbic acid and glutamate levels had increased, dehydroascorbic acid and GSH decreased, whereas catabolites of high-energy phosphates (inosine, hypoxanthine, xanthine, and uric acid) were unchanged. In addition, gliosis was observed in both striatum and substantia nigra compacta (SNc). Sections of SNc showed some terminal deoxynucleotidyl transferase-mediated 2'-deoxyuridine 5'-triphosphate nick end labeling (TUNEL)-positive cells. Neurochemical parameters of dopaminergic activity showed a trend toward recovery 3 days after MPTP discontinuance. At this time point, TUNEL-positive cells were detected in SNc; some of them showed nuclei with neuronal morphology. A late (days 6-11) increase in striatal dopamine oxidative metabolism, ascorbic acid oxidative status, and catabolites of high-energy phosphates were observed concomitant with nigral neuron and nigrostriatal glial cell apoptotic death, as revealed by TUNEL, acridine orange, and Hoechst staining, and transmission electron microscopy. These data suggest that MPTP-induced activation/apoptotic death of glial cells plays a key role in the sequential linkage of neurochemical and cellular events leading to dopaminergic nigral neuron apoptotic death

Metabolomic analysis of mouse prefrontal cortex reveals upregulated analytes during wakefulness compared to sleep

By identifying endogenous molecules in brain extracellular fluid metabolomics can provide insight into the regulatory mechanisms and functions of sleep. Here we studied how the cortical metabolome changes during sleep, sleep deprivation and spontaneous wakefulness. Mice were implanted with electrodes for chronic sleep/wake recording and with microdialysis probes targeting prefrontal and primary motor cortex. Metabolites were measured using ultra performance liquid chromatography-high resolution mass spectrometry. Sleep/wake changes in metabolites were evaluated using partial least squares discriminant analysis, linear mixed effects model analysis of variance, and machine-learning algorithms. More than 30 known metabolites were reliably detected in most samples. When used by a logistic regression classifier, the profile of these metabolites across sleep, spontaneous wake, and enforced wake was sufficient to assign mice to their correct experimental group (pair-wise) in 80–100% of cases. Eleven of these metabolites showed significantly higher levels in awake than in sleeping mice. Some changes extend previous findings (glutamate, homovanillic acid, lactate, pyruvate, tryptophan, uridine), while others are novel (D-gluconate, N-acetyl-beta-alanine, N-acetylglutamine, orotate, succinate/methylmalonate). The upregulation of the de novo pyrimidine pathway, gluconate shunt and aerobic glycolysis may reflect a wake-dependent need to promote the synthesis of many essential components, from nucleic acids to synaptic membranes

Brodifacoum Levels and Biomarkers in Coastal Fish Species following a Rodent Eradication in an Italian Marine Protected Area: Preliminary Results

Brodifacoum is the most common rodenticide used for the eradication of invasive rodents from islands. It blocks the vitamin K cycle, resulting in hemorrhages in target mammals. Non-target species may be incidentally exposed to brodifacoum, including marine species. A case study conducted on the Italian Marine Protected Area of Tavolara Island was reported after a rodent eradication using the aerial broadcast of a brodifacoum pellet. Brodifacoum presence and effects on non-target marine organisms were investigated. Different fish species were sampled, and a set of analyses was conducted to determine vitamin K and vitamin K epoxide reductase concentrations, prothrombin time, and erythrocytic nuclear abnormalities (ENA) assay. In all the examined organisms, brodifacoum was not detected. The results obtained showed differences in vitamin K and vitamin K epoxide concentrations among the samples studied, with a positive correlation for three species between vitamin K, vitamin K epoxide, and fish weight. The prothrombin time assay showed a good blood clotting capacity in the fish. Higher abnormality values were recorded for four species. The results of this study suggest that it is possible to hypothesize that the sampled fish were not likely to have been exposed to brodifacoum and that consequently there are no negative issues concerning human consumption

Ultrastructural effects of sleep and wake on the parallel fiber synapses of the cerebellum

Multiple evidence in rodents shows that the strength of excitatory synapses in the cerebral cortex and hippocampus is greater after wake than after sleep. The widespread synaptic weakening afforded by sleep is believed to keep the cost of synaptic activity under control, promote memory consolidation, and prevent synaptic saturation, thus preserving the brain's ability to learn day after day. The cerebellum is highly plastic and the Purkinje cells, the sole output neurons of the cerebellar cortex, are endowed with a staggering number of excitatory parallel fiber synapses. However, whether these synapses are affected by sleep and wake is unknown. Here, we used serial block face scanning electron microscopy to obtain the full 3D reconstruction of more than 7000 spines and their parallel fiber synapses in the mouse posterior vermis. This analysis was done in mice whose cortical and hippocampal synapses were previously measured, revealing that average synaptic size was lower after sleep compared to wake with no major changes in synapse number. Here, instead, we find that while the average size of parallel fiber synapses does not change, the number of branched synapses is reduced in half after sleep compared to after wake, corresponding to similar to 16% of all spines after wake and similar to 8% after sleep. Branched synapses are harbored by two or more spines sharing the same neck and, as also shown here, are almost always contacted by different parallel fibers. These findings suggest that during wake, coincidences of firing over parallel fibers may translate into the formation of synapses converging on the same branched spine, which may be especially effective in driving Purkinje cells to fire. By contrast, sleep may promote the off-line pruning of branched synapses that were formed due to spurious coincidences

Chemico-biological characterization of Torpedino Di Fondi® tomato fruits. A comparison with San Marzano cultivar at two ripeness stages

Torpedino di Fondi (TF) is a hybrid tomato landrace developed in Sicily and recently introduced in the south Lazio area along with the classical San Marzano (SM) cultivar. The present study aimed at characterizing TF tomatoes at both pink and red ripening stages, and at comparing them with traditional SM tomatoes. A multidisciplinary approach consisting of morphological, chemical (FT‐ICR MS, NMR, HPLC, and spectrophotometric methods), and biological (antioxidant and antifungal in vitro activity) analyses was applied. Morphological analysis confirmed the mini‐ San Marzano nature and the peculiar crunchy and solid consistency of TF fruits. Pink TF tomatoes displayed the highest content of hydrophilic antioxidants, like total polyphenols (0.192 mg/g), tannins (0.013 mg/g), flavonoids (0.204 mg/g), and chlorophylls a (0.344 mg/g) and b (0.161 mg/g), whereas red TF fruits were characterized by the highest levels of fructose (3000 mg/100 g), glucose (2000 mg/100 g), tryptophan (2.7 mg/100 g), phenylalanine (13 mg/100 g), alanine (25 mg/100 g), and total tri‐unsaturated fatty acids (13% mol). Red SM fruits revealed the greatest content of lipophilic antioxidants, with 1234 mg/g of total carotenoids. In agreement with phenolics content, TF cultivar showed the greatest antioxidant activity. Lastly, red TF inhibited Candida species (albicans, glabrata and krusei) growth

Ropinirole and pramipexole promote structural plasticity in human iPSC- derived dopaminergic neurons via BDNF and mTOR signaling

The antiparkinsonian ropinirole and pramipexole are D3 receptor- (D3R-) preferring dopaminergic (DA) agonists used as adjunctive therapeutics for the treatment resistant depression (TRD). While the exact antidepressant mechanism of action remains uncertain, a role for D3R in the restoration of impaired neuroplasticity occurring in TRD has been proposed. Since D3R agonists are highly expressed on DA neurons in humans, we studied the effect of ropinirole and pramipexole on structural plasticity using a translational model of human-inducible pluripotent stem cells (hiPSCs). Two hiPSC clones from healthy donors were differentiated into midbrain DA neurons. Ropinirole and pramipexole produced dose-dependent increases of dendritic arborization and soma size after 3 days of culture, effects antagonized by the selective D3R antagonists SB277011-A and S33084 and by the mTOR pathway kinase inhibitors LY294002 and rapamycin. All treatments were also effective in attenuating the D3R-dependent increase of p70S6-kinase phosphorylation. Immunoneutralisation of BDNF, inhibition of TrkB receptors, and blockade of MEK-ERK signaling likewise prevented ropinirole-induced structural plasticity, suggesting a critical interaction between BDNF and D3R signaling pathways. The highly similar profiles of data acquired with DA neurons derived from two hiPSC clones underpin their reliability for characterization of pharmacological agents acting via dopaminergic mechanisms