Microelectrode arrays of diamond-insulated graphitic channels for real time detection of exocytotic events from cultured chromaffin cells and slices of adrenal glands

A microstructured graphitic 4x4 multielectrode array was embedded in a single crystal diamond substrate (4x4 {uG-SCD MEA) for real-time monitoring of exocytotic events from cultured chromaffin cells and adrenal slices. The current approach relies on the development of a parallel ion beam lithographic technique, which assures the time effective fabrication of extended arrays with reproducible electrode dimensions. The reported device is suitable for performing amperometric and voltammetric recordings with high sensitivity and temporal resolution, by simultaneously acquiring data from 16 rectangularly shaped microelectrodes (20x3.5 um^2) separated by 200 um gaps. Taking advantage of the array geometry we addressed the following specific issues: i) detect both the spontaneous and KCl-evoked secretion simultaneously from several chromaffin cells directly cultured on the device surface, ii) resolve the waveform of different subsets of exocytotic events, iii) monitoring quantal secretory events from thin slices of the adrenal gland. The frequency of spontaneous release was low (0.12 Hz and 0.3 Hz respectively for adrenal slices and cultured cells) and increased up to 0.9 Hz after stimulation with 30 mM KCl in cultured cells. The spike amplitude as well as rise and decay time were comparable with those measured by carbon fiber microelectrodes and allowed to identify three different subsets of secretory events associated to "full fusion" events, "kiss and-run" and "kiss-and-stay" exocytosis, confirming that the device has adequate sensitivity and time resolution for real-time recordings. The device offers the significant advantage of shortening the time to collect data by allowing simultaneous recordings from cell populations either in primary cell cultures or in intact tissues

Kelvin probe characterization of buried graphitic microchannels in single-crystal diamond

In this work, we present an investigation by Kelvin Probe Microscopy (KPM) of buried graphitic microchannels fabricated in single-crystal diamond by direct MeV ion microbeam writing. Metal deposition of variable-thickness masks was adopted to implant channels with emerging endpoints and high temperature annealing was performed in order to induce the graphitization of the highly-damaged buried region. When an electrical current was flowing through the biased buried channel, the structure was clearly evidenced by KPM maps of the electrical potential of the surface region overlying the channel at increasing distances from the grounded electrode. The KPM profiling shows regions of opposite contrast located at different distances from the endpoints of the channel. This effect is attributed to the different electrical conduction properties of the surface and of the buried graphitic layer. The model adopted to interpret these KPM maps and profiles proved to be suitable for the electronic characterization of buried conductive channels, providing a non-invasive method to measure the local resistivity with a micrometer resolution. The results demonstrate the potential of the technique as a powerful diagnostic tool to monitor the functionality of all-carbon graphite/diamond devices to be fabricated by MeV ion beam lithography.Comment: 21 pages, 5 figure

Mid Campanian-Lower Maastrichtian magnetostratigraphy of the James Ross Basin, Antarctica: Chronostratigraphical implications

The James Ross Basin, in the northern Antarctic Peninsula, exposes which is probably the world thickest and most complete Late Cretaceous sedimentary succession of southern high latitudes. Despite its very good exposures and varied and abundant fossil fauna, precise chronological determination of its infill is still lacking. We report results from a magnetostratigraphic study on shelfal sedimentary rocks of the Marambio Group, southeastern James Ross Basin, Antarctica. The succession studied covers a ~1,200 m-thick stratigraphic interval within the Hamilton Point, Sanctuary Cliffs and Karlsen Cliffs Members of the Snow Hill Island Formation, the Haslum Crag Formation, and the lower López de Bertodano Formation. The basic chronological reference framework is given by ammonite assemblages, which indicate a Late Campanian – Early Maastrichtian age for the studied units. Magnetostratigraphic samples were obtained from five partial sections located on James Ross and Snow Hill islands, the results from which agree partially with this previous biostratigraphical framework. Seven geomagnetic polarity reversals are identified in this work, allowing to identify the Chron C32/C33 boundary in Ammonite Assemblage 8-1, confirming the Late Campanian age of the Hamilton Point Member. However, the identification of the Chron C32/C31 boundary in Ammonite Assemblage 8-2 assigns the base of the Sanctuary Cliffs Member to the early Maastrichtian, which differs from the Late Campanian age previously assigned by ammonite biostratigraphy. This magnetostratigraphy spans ~14 Ma of sedimentary succession and together with previous partial magnetostratigraphies on Early-Mid Campanian and Middle Maastrichtian to Danian columns permits a complete and continuous record of the Late Cretaceous distal deposits of the James Ross Basin. This provides the required chronological resolution to solve the intra-basin and global correlation problems of the Late Cretaceous in the Southern Hemisphere in general and in the Weddellian province in particular, given by endemism and diachronic extinctions on invertebrate fossils, including ammonites. The new chronostratigraphic scheme allowed us to calculate sediment accumulation rates for almost the entire Late Cretaceous infill of the distal James Ross Basin (the Marambio Group), showing a monotonous accumulation for more than 8 Myr during the upper Campanian and a dramatic increase during the early Maastrichtian, controlled by tectonic and/or eustatic causes.Fil: Milanese, Florencia Nidia. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Geociencias Básicas, Aplicadas y Ambientales de Buenos Aires. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Geociencias Básicas, Aplicadas y Ambientales de Buenos Aires; ArgentinaFil: Olivero, Eduardo Bernardo. Universidad Nacional de Tierra del Fuego; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Austral de Investigaciones Científicas; ArgentinaFil: Raffi, María Eugenia. Universidad Nacional de Tierra del Fuego; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Austral de Investigaciones Científicas; ArgentinaFil: Franceschinis, Pablo Reinaldo. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Geociencias Básicas, Aplicadas y Ambientales de Buenos Aires. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Geociencias Básicas, Aplicadas y Ambientales de Buenos Aires; ArgentinaFil: Gallo, Leandro César. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Geociencias Básicas, Aplicadas y Ambientales de Buenos Aires. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Geociencias Básicas, Aplicadas y Ambientales de Buenos Aires; ArgentinaFil: Skinner, Steven M.. California State University; Estados UnidosFil: Mitchell, Ross N.. California Institute of Technology; Estados UnidosFil: Kirschvink, Joseph L.. California Institute of Technology; Estados Unidos. Tokyo Institute of Technology; JapónFil: Rapalini, Augusto Ernesto. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Geociencias Básicas, Aplicadas y Ambientales de Buenos Aires. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Geociencias Básicas, Aplicadas y Ambientales de Buenos Aires; Argentin

A methodological proposal and tool support for the HL7 standards compliance in the development of health information systems

Health information systems are increasingly complex, and their development is presented as a challenge for software development companies offering quality, maintainable and interoperable products. HL7 (Health level 7) International, an international non-profit organization, defines and maintains standards related to health information systems. However, the modelling languages proposed by HL7 are far removed from standard languages and widely known by software engineers. In these lines, NDT is a software development methodology that has a support tool called NDT-Suite and is based, on the one hand, on the paradigm of model-driven engineering and, on the other hand, in UML that is a widely recognized standard language. This paper proposes an extension of the NDT methodology called MoDHE (Model Driven Health Engineering) to offer software engineers a methodology capable of modelling health information systems conforming to HL7 using UML domain models

All-carbon multi-electrode array for real-time in vitro measurements of oxidizable neurotransmitters

We report on the ion beam fabrication of all-carbon multi electrode arrays (MEAs) based on 16 graphitic micro-channels embedded in single-crystal diamond (SCD) substrates. The fabricated SCD-MEAs are systematically employed for the in vitro simultaneous amperometric detection of the secretory activity from populations of chromaffin cells, demonstrating a new sensing approach with respect to standard techniques. The biochemical stability and biocompatibility of the SCD-based device combined with the parallel recording of multi-electrodes array allow: i) a significant time saving in data collection during drug screening and/or pharmacological tests over a large number of cells, ii) the possibility of comparing altered cell functionality among cell populations, and iii) the repeatition of acquisition runs over many cycles with a fully non-toxic and chemically robust bio-sensitive substrate.Comment: 24 pages, 5 figure

fMRI in patients with lumbar disc disease: a paradigm to study patients over time

Low back pain is a common human ailment. It is estimated that over 70% of the population will experience low back pain that will require medication and/or medical attention. There are many causes for low back pain, one being herniation of the discs of the lumbar spine. Treatment options are very limited. Why patients develop chronic pain especially when there is no known organic cause or when the offending painful stimulus has been removed remains poorly understood. Functional magnetic resonance imaging (fMRI) is a technique that allows researchers to image which regions of the brain that are activated during motor, cognitive, and sensory experiences. Using fMRI to study pain has revealed new information about how the brain responds to painful stimuli and what regions of the brain are activated during pain. However, many of the paradigms used do not replicate the subject’s pain or use painful stimuli in volunteers without pain. Also, following patients from their acute phase of pain to the chronic phase with serial fMRI has not been performed. In this study we developed a paradigm that would allow studying patients with low back pain and leg pain including lumbar radiculopathy to better mimic a clinical pain syndrome and to have a method of following patients with this type of pain over time