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

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

The efficiency of immobilised glutamate oxidase decreases with surface enzyme loading: an electrostatic effect, and reversal by a polycation significantly enhances biosensor sensitivity

The apparent Michaelis constant, KM, for glutamate oxidase (GluOx) immobilised on Pt electrodes increased systematically with enzyme loading. The effect was due, at least in part, to electrostatic repulsion between neighbouring oxidase molecules and the anionic substrate, glutamate (Glu). This understanding has allowed us to increase the Glu sensitivity of GluOx-based amperometric biosensors in the linear response region (100 ± 11 nA cmâ2µMâ1 at pH 7.4; SD, n = 23) by incorporating a polycation (polyethyleneimine, PEI) to counterbalance the polyanionic protein. Differences in the behaviour of glucose biosensors of a similar configuration highlight a limitation of using glucose oxidase as a model enzyme in biosensor design

Control of the Oxygen Dependence of an Implantable Polymer/Enzyme Composite Biosensor for Glutamate

Biosensors for glutamate (Glu) were fabricated from Teflon-coated Pt wire (cylinders and disks), modified with the enzyme glutamate oxidase (GluOx) and electrosynthesized polymer PPD, poly(o-phenylenediamine). The polymer/enzyme layer was deposited in two configurations:  enzyme before polymer (GluOx/PPD) and enzyme after polymer (PPD/GluOx). These four biosensor designs were characterized in terms of response time, limit of detection, Michaelis−Menten parameters for Glu (Jmax and KM(Glu)), sensitivity to Glu in the linear response region, and dependence on oxygen concentration, KM(O2). Analysis showed that the two polymer/enzyme configurations behaved similarly on both cylinders and disks. Although the two geometries showed different behaviors, these differences could be explained in terms of higher enzyme loading density on the disks; in many analyses, the four designs behaved like a single population with a range of GluOx loading. Enzyme loading was the key to controlling the KM(O2) values of these first generation biosensors. The counterintuitive, and beneficial, behavior that biosensors with higher GluOx loading displayed a lower oxygen dependence was explained in terms of the effects of enzyme loading on the affinity of GluOx for its anionic substrate. Some differences between the properties of surface immobilized GluOx and glucose oxidase are highlighted

The efficiency of immobilised glutamate oxidase decreases with surface enzyme loading: an electrostatic effect, and reversal by a polycation significantly enhances biosensor sensitivity

The apparent Michaelis constant, KM, for glutamate oxidase (GluOx) immobilised on Pt electrodes increased systematically with enzyme loading. The effect was due, at least in part, to electrostatic repulsion between neighbouring oxidase molecules and the anionic substrate, glutamate (Glu). This understanding has allowed us to increase the Glu sensitivity of GluOx-based amperometric biosensors in the linear response region (100 ± 11 nA cmâ2µMâ1 at pH 7.4; SD, n = 23) by incorporating a polycation (polyethyleneimine, PEI) to counterbalance the polyanionic protein. Differences in the behaviour of glucose biosensors of a similar configuration highlight a limitation of using glucose oxidase as a model enzyme in biosensor design

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

Neuroprotective effect of (R)-(-)-linalool on oxidative stress in PC12 cells

Background: Oxidative stress plays an important role in neurodegeneration, pain and inflammation. (R)-(-)- linalool (LIN) is endowed with neuroprotective, anti-nociceptive and anti-inflammatory properties. Purpose: The present study aims at investigating the hypothesis that LIN’s neuroprotective, antinociceptive and anti-inflammatory properties descend from its ability to act as antioxidant. The study challenges this hypothesis by verifying whether LIN may counteract hydrogen peroxide (H 2 O 2 )-induced oxidative stress in PC12 cells. Methods: In H 2 O 2 -exposed PC12 cells, LIN was tested on a) cell viability, measured by 3-(4,5-dimethylthiazol-2- yl)-2,5-diphenyltetrazolium bromide (MTT), b) damage of plasma membrane, measured by lactate dehydrogenase (LDH) release, c) intracellular levels of reactive-oxygen-species (ROS), d) apoptosis and e) cell cycle distribution. Results: Under H 2 O 2 -induced cell viability reduction, LIN protects PC12 cells. Likewise, LIN protects cells from oxidative damage by preventing the H 2 O 2 -dependent increase of LDH release, counteracts intracellular ROS overproduction and reduces H 2 O 2 -induced apoptosis. Finally, the results of the cell cycle analysis from cells exposed to H 2 O 2 indicate that LIN incubation reduces the number of cells induced into quiescence by H 2 O 2 in the G2/M phase. Conclusions: These findings indicate that LIN protects PC12 cells from H 2 O 2 -induced oxidative stress. This mech- anism could justify the neuroprotective, anti-nociceptive and anti-inflammatory effects of this compound and suggest LIN as a potential therapeutic agent for the management oxidative stress-mediated pain

New perspective for an old drug: Can naloxone be considered an antioxidant agent?

Background: Experimental evidence indicates that Naloxone (NLX) holds antioxidant properties. The present study aims at verifying the hypothesis that NLX could prevent oxidative stress induced by hydrogen peroxide (H2O2) in PC12 cells.Methods: To investigate the antioxidant effect of NLX, initially, we performed electrochemical experiments by means of platinum-based sensors in a cell-free system. Subsequently, NLX was tested in PC12 cells on H2O2induced overproduction of intracellular levels of reactive-oxygen-species (ROS), apoptosis, modification of cells' cycle distribution and damage of cells' plasma membrane.Results: This study reveals that NLX counteracts intracellular ROS production, reduces H2O2-induced apoptosis levels, and prevents the oxidative damage-dependent increases of the percentage of cells in G2/M phase. Likewise, NLX protects PC12 cells from H2O2- induced oxidative damage, by preventing the lactate dehydrogenase (LDH) release. Moreover, electrochemical experiments confirmed the antioxidant properties of NLX.Conclusion: Overall, these findings provide a starting point for studying further the protective effects of NLX on oxidative stress

Real-Time Monitoring of Brain Tissue Oxygen Using a Miniaturized Biotelemetric Device Implanted in Freely Moving Rats

A miniaturized biotelemetric device for the amperometric detection of brain tissue oxygen is presented. The new system, derived from a previous design, has been coupled with a carbon microsensor for the real-time detection of dissolved O(2) in the striatum of freely moving rats. The implantable device consists of a single-supply sensor driver, a current-to-voltage converter, a microcontroller, and a miniaturized data transmitter. The oxygen current is converted to a digital value by means of an analog-to-digital converter integrated in a peripheral interface controller (PIC). The digital data is sent to a personal computer using a six-byte packet protocol by means of a miniaturized 434 MHz amplitude modulation (AM) transmitter. The receiver unit is connected to a personal computer (PC) via a universal serial bus. Custom developed software allows the PC to store and plot received data. The electronics were calibrated and tested in vitro under different experimental conditions and exhibited high stability, low power consumption, and good linear response in the nanoampere current range. The in vivo results confirmed previously published observations on oxygen dynamics in the striatum of freely moving rats. The system serves as a rapid and reliable model for studying the effects of different drugs on brain oxygen and brain blood flow and it is suited to work with direct-reduction sensors or O(2)-consuming biosensors