Direct monitoring of ethanol in the brain

Introduction In the past few decades, ethanol has assumed the role of the most widespread psychotropic agent in Western society because of its availability to the youth and adults and also because it is generally considered legal in many societies. It is known that the alcohol can have significant relapses on the central nervous system; hence, there is a need for monitoring the toxicokinetics and the effects of ethanol on the brain with the most appropriate techniques. Among the techniques that aim to measure ethanol concentration in the brain, microdialysis has been the most widely used, but because of its invasiveness, associated with low temporal resolution, and the necessity of using connecting tubes to carry out the experiments, it is not particularly suitable for clinical trials. Recently, electrochemical biosensors, also minimally invasive, have been developed, which offer the possibility of monitoring the real-time variations of ethanol concentrations in the brain of animal models due to the very small dimensions of the transducer electrode. Recently, non-invasive methods have been used for the direct monitoring of alcohol in the brain, which use spectroscopic techniques such as magnetic resonance spectroscopy and magnetic resonance imaging or positron emission tomography, which are principally used to monitor ethanol metabolites. The aim of this review is to discuss all the techniques used to monitor brain ethanol and highlight their strengths and weaknesses. Conclusion Microdialysis and biosensors are primarily used in preclinical studies; both are very reliable techniques, but for invasiveness, they can only be used in animal models. Alternatively, spectroscopic techniques are suitable for both preclinical and clinical studies, and are not exclusive for animal models.</br

Design and construction of a distributed sensor NET for biotelemetric monitoring of brain energetic metabolism using microsensors and biosensors

Neurochemical pathways involved in brain physiology or disease pathogenesis are mostly unknown either in physiological conditions or in neurodegenerative diseases. Nowadays the most frequent usage for biotelemetry is in medicine, in cardiac care units or step-down units in hospitals, even if virtually any physiological signal could be transmitted (FCC, 2000; Leuher, 1983; Zhou et al., 2002). In this chapter we present a wireless device connected with microsensors and biosensors capable to detect real-time variations in concentrations of important compounds present in central nervous system (CNS) and implicated in brain energetic metabolism (Bazzu et al., 2009; Calia et al., 2009)

Synthesis and study of polyhydroxylated phenol derivatives with potential cosmetic and phytoiatric applications

Tyrosinase (polyphenol oxidase, E.C. 1.14.18.1) and laccase (phenol oxidase, E.C. 1.10.3.2) are multifunctional copper-containing enzymes, that are keys in melanin biosynthesis, melanisation in animals and browning in plants. Our study is aimed to prepare new monomer and dimer phenol derivatives as potential inhibitors of melanin production starting from natural hydroxylated aromatic units

Selective and sensitive poly-<i>ortho</i>-phenylenediamine-shielded microsensore and biosensors for in vivo neurochemical monitoring

Different methodologies are being developed, such as imaging, spectroscopy and electrochemistry, to study neurochemical dynamics in cell cultures or in intact brain [1-2]. One of these techniques involves the in-situ detection of biologically active molecules, including nitric oxide (NO) [3], glucose [4], glutamate (GLUT) [5-6] and lactate [1,7], in brain extracellular fluid (ECF), using implanted microsensors and biosensors. NO is a water-soluble free radical that readily diffuses through membranes and its actions in the CNS are largely studied

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

Neurological morphofunctional differentiation induced by REAC technology in PC12: a neuro protective model for Parkinson's disease

Research for the use of physical means, in order to induce cell differentiation for new therapeutic strategies, is one of the most interesting challenges in the field of regenerative medicine, and then in the treatment of neurodegenerative diseases, Parkinson’s disease (PD) included. The aim of this work is to verify the effect of the radio electric asymmetric conveyer (REAC) technology on the PC12 rat adrenal pheochromocytoma cell line, as they display metabolic features of PD. PC12 cells were cultured with a REAC regenerative tissue optimization treatment (TO-RGN) for a period ranging between 24 and 192 hours. Gene expression analysis of specific neurogenic genes, as neurogenin-1, beta3-tubulin and Nerve growth factor, together with the immunostaining analysis of the specific neuronal protein beta3-tubulin and tyrosine hydroxylase, shows that the number of cells committed toward the neurogenic phenotype was significantly higher in REAC treated cultures, as compared to control untreated cells. Moreover, MTT and Trypan blue proliferation assays highlighted that cell proliferation was significantly reduced in REAC TO-RGN treated cells. These results open new perspectives in neurodegenerative diseases treatment, particularly in PD. Further studies will be needed to better address the therapeutic potential of the REAC technology

Modifications of poly(o-phenylenediamine) permselective layer on Pt-Ir for biosensor application in neurochemical monitoring

Reports that globular proteins could enhance the interference blocking ability of the PPD (poly(o-phenylenediamine) layer used as a permselective barrier in biosensor design, prompted this study where a variety of modifying agents were incorporated into PPD during its electrosynthesis on Pt-Ir electrodes. Trapped molecules, including fibrous proteins and β-cyclodextrin, altered the polymer/modifier composite selectivity by affecting the sensitivity to both H2O2 (signal molecule in many enzyme-based biosensors) and the archetypal interference species, ascorbic acid. A comparison of electrochemical properties of Pt and a Pt-Ir alloy suggests that the benefits of the latter, more rigid, metal can be exploited in PPD-based biosensor design without significant loss of backward compatibility with studies involving pure Pt

Designing sensitive and selective polymer/enzyme composite biosensors for brain monitoring <i>in vivo</i>

Amperometric polymer/enzyme composite (PEC) biosensors, incorporating a poly(o-phenylenediamine) ultra-thin permselective barrier, possess a variety of characteristics that make them suitable for monitoring brain energy and neurotransmitter dynamics in vivo. This review highlights PEC sensitivity and selectivity parameters, which allow development of the basic design in a systematic way in order to improve their performance and to diversify the analyte range of these novel probes of brain function

Endogenous melatonin protects L-DOPA from autoxidation in the striatal extracellular compartment of the freely moving rat: potential implication for long-term L-DOPA therapy in Parkinson's disease

We previously showed, using microdialysis, that autoxidation of exogenous L-dihydroxyphenylalanine (L-DOPA) occurs in vivo in the extracellular compartment of the freely moving rat, with a consequent formation of L-DOPA semiquinone (L-DOPA-SQ). In the present study, intrastriatal infusion of L-DOPA (1.0 μm for 200 min) increased dialysate L-DOPA concentrations (maximum increases up to 116-fold baseline values); moreover, L-DOPA-SQ was detected in dialysates. Individual dialysate concentrations of L-DOPA were negatively correlated with those of L-DOPA-SQ. Co-infusion of N-acetylcysteine (100 μm) or melatonin (50 μm) increased L-DOPA (up to 151- and 246-fold, respectively) and decreased L-DOPA-SQ (by about 53% and 87%, respectively) dialysate concentrations. Systemic L-DOPA [25 mg/kg intraperitoneally (i.p.) twice in a 12-h interval] significantly increased striatal baseline dialysate concentrations of L-DOPA and decreased dopamine (DA) and ascorbic acid (AsAc) concentrations, when compared with controls. Following systemic L-DOPA, L-DOPA-SQ was detected in dialysates. Endogenous melatonin was depleted in rats maintained on a 24-h light cycle for 1 wk. In melatonin-depleted rats, systemic L-DOPA induced a smaller increase in dialysate L-DOPA, a greater increase in L-DOPA-SQ formation, and a greater reduction in DA and AsAc dialysate concentrations. Co-administration of melatonin (5.0 mg/kg, i.p., twice in a 12-h interval) with L-DOPA, in control as well as in light-exposed rats, significantly increased dialysate L-DOPA concentrations, greatly inhibited L-DOPA-SQ formation, and restored up to the control values dialysate DA and AsAc concentrations. These findings demonstrate that endogenous melatonin protects exogenous L-DOPA from autoxidation in the extracellular compartment of the striatum of freely moving rats; moreover, systemic co-administration of melatonin with L-DOPA markedly increases striatal L-DOPA bioavailability in control as well as in melatonin-depleted rats. These results may be of relevance to the long-term L-DOPA therapy of Parkinson's disease

Signalling pathways in the nitric oxide donor-induced dopamine release in the striatum of freely moving rats: evidence that exogenous nitric oxide promotes Ca²⁺ entry through store-operated channels

We showed previously, using in vitro microdialysis, that the activation of the soluble guanylate cyclase (sGC)/cyclic GMP pathway was the underlying mechanism of the extracellular Ca2+-dependent effects of exogenous NO on dopamine (DA) secretion from PC12 cells. In this study, the co-infusion of the sGC inhibitor 1H-[1,2,4]oxadiazolo[4,3] quinoxalin-1-one (ODQ) failed to affect the NO donor 3- morpholinosydnonimine (SIN-1, 5.0 mM)-induced DA increase (sevenfold baseline) in dialysates from the striatum of freely moving rats. Ca2+ omission from the perfusion fluid abolished baseline DA release but did not affect SIN-1-induced DA increases. The reintroduction of Ca2+ in the perfusion fluid restored the baseline dialysate DA; however, when Ca2+ reintroduction was associated with the infusion of either SIN-1 or the NO-donor S-nitrosoglutathione (SNOG), a sustained DA overflow was observed. DA overflow was selectively inhibited by the co-infusion of the store-operated channel blocker 2-aminoethoxydiphenyl borate. The chelation of intracellular Ca2+ by co-infusing 1,2-bis (o-amino-phenoxy)ethane-N,N,N’,N’-tetraacetic acid tetra (acetoxymethyl) ester (BAPTA-AM, 0.2 mM) greatly potentiated both SIN-1- and SNOG-induced increases in dialysate DA. BAPTA-AM-induced potentiation was inhibited by Ca2+ omission. We conclude that the sGC/ cyclic GMP pathway is not involved in the extracellular Ca2+-independent exogenous NO-induced striatal DA release; however, when intracellular Ca2+ is either depleted (by Ca2+ omission) or chelated (by BAPTA-AM co-infusion), exogenous NO does promote Ca2+ entry, most likely through store-operated channels, with a consequent further increase in DA release