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

Correlations in quantum thermodynamics: Heat, work, and entropy production

We provide a characterization of energy in the form of exchanged heat and work between two interacting constituents of a closed, bipartite, correlated quantum system. By defining a binding energy we derive a consistent quantum formulation of the first law of thermodynamics, in which the role of correlations becomes evident, and this formulation reduces to the standard classical picture in relevant systems. We next discuss the emergence of the second law of thermodynamics under certain---but fairly general---conditions such as the Markovian assumption. We illustrate the role of correlations and interactions in thermodynamics through two examples.Comment: 16 page

Nanodiamonds-induced effects on neuronal firing of mouse hippocampal microcircuits

Fluorescent nanodiamonds (FND) are carbon-based nanomaterials that can efficiently incorporate optically active photoluminescent centers such as the nitrogen-vacancy complex, thus making them promising candidates as optical biolabels and drug-delivery agents. FNDs exhibit bright fluorescence without photobleaching combined with high uptake rate and low cytotoxicity. Focusing on FNDs interference with neuronal function, here we examined their effect on cultured hippocampal neurons, monitoring the whole network development as well as the electrophysiological properties of single neurons. We observed that FNDs drastically decreased the frequency of inhibitory (from 1.81 Hz to 0.86 Hz) and excitatory (from 1.61 Hz to 0.68 Hz) miniature postsynaptic currents, and consistently reduced action potential (AP) firing frequency (by 36%), as measured by microelectrode arrays. On the contrary, bursts synchronization was preserved, as well as the amplitude of spontaneous inhibitory and excitatory events. Current-clamp recordings revealed that the ratio of neurons responding with AP trains of high-frequency (fast-spiking) versus neurons responding with trains of low-frequency (slow-spiking) was unaltered, suggesting that FNDs exerted a comparable action on neuronal subpopulations. At the single cell level, rapid onset of the somatic AP ("kink") was drastically reduced in FND-treated neurons, suggesting a reduced contribution of axonal and dendritic components while preserving neuronal excitability.Comment: 34 pages, 9 figure

Macro Minerals and Trace Elements in Milk of Dairy Buffaloes and Cows Reared in Mediterranean Areas

Aim of this study was to evaluate the differences in Ca, P, K, Na, Mg, Zn, Fe, Cu, Mn, Se, Mo, Co, Li, B, Ti, Rb, and Sr concentrations in milk from buffaloes and cows reared in the same farm in Mediterranean areas and fed diets including the same ingredients. Individual milk samples were obtained from 32 Mediterranean buffaloes and 29 Italian Friesian cows and samples of milk, dietary ingredients and drinking water were analyzed for the investigated chemical elements by inductively coupled plasma-mass spectrometry. Data about milk element concentrations were processed by one-way analysis of variance. Buffalo milk contains higher concentrations of Ca, P, Mg, Zn, Fe, Cu, B, Ti, and Sr, and lower concentrations of K, Na, Mo, Li, and Rb compared to cow milk, whereas milk from both species contains similar concentrations of Mn, Se, and Co. The concentrations of the investigated elements in the diet were similar for both species and the differences observed between buffalo and cow milk were not dependent on environmental factors

Distribution of selected trace elements in the major fractions of donkey milk

The aim of this study was to evaluate the concentrations of Zn, Cu, Mn, Se, Mo, Co, Li, B, Ti, Cr, Rb, Sr, Cd, and Pb in donkey milk and their distribution in major milk fractions (i.e., fat, casein, whey proteins, and aqueous phase). Individual milk samples were provided by 16 clinically healthy lactating donkeys. Subsequent centrifugation, ultracentrifugation, and ultrafiltration were carried out to remove fat, casein, and whey proteins to obtain skim milk, a supernatant whey fraction, and the aqueous phase of milk, respectively. Concentrations of the elements were measured in whole milk and fractions by inductively coupled plasma-mass spectrometry, and the concentrations associated with fat, casein, and whey proteins were then calculated. The effect of removal of fat, casein, and whey proteins was determined by repeated-measures ANOVA. The fat fraction of donkey milk carried a small (∼4.5% to 13.5%) but significant proportion of Mo, Co, Ti, Cr, and Sr. The casein fraction in donkey milk carried almost all milk Zn, a majority of Cu and Mn, and most of Mo, Ti, and Sr. Relevant proportions, between 20% and 36%, of Se, Co, and Cr were also associated with caseins. The majority of Se, Co, Li, B, Cr, and Rb, and relevant proportions of Mn, Mo, Ti, and Sr were found in soluble form (ultracentrifuged samples) and distributed between whey proteins and the aqueous phase of milk (ultrafiltered samples). Whey proteins in donkey milk carried the majority of milk Se and Co. All Li and B was present in the aqueous phase of milk, which also contained most Rb and Cr, and 17% to 42% of Mn, Se, Mo, Co, Ti, and Sr

Initial investigation of athletes’ electrocardiograms acquired by wearable sensors during the pre-exercise phase

Aim: The aim of this study is to support large-scale prevention programs fighting sport-related sudden cardiac death by providing a set of electrocardiographic features representing a starting point in the development of normal reference values for the pre-exercise phase. Background: In people with underlying, often unknown, cardiovascular abnormalities, increased cardiovascular load during exercise can trigger sport-related sudden cardiac death. Prevention remains the only weapon to contrast sport-related sudden cardiac death. So far, no reference values have been proposed for electrocardiograms of athletes acquired with wearable sensors in the pre-exercise phase, consisting of the few minutes immediately before the beginning of the training session. Objective: To perform an initial investigation of athletes’ electrocardiograms acquired by wearable sensors during the pre-exercise phase. Methods: The analyzed electrocardiograms, acquired through BioHarness 3.0 by Zephyr, belong to 51 athletes (Sport Database and Cycling Database of the Cardiovascular Bioengineering Lab of the Università Politecnica delle Marche, Italy). Preliminary values consist of interquartile ranges of six electrocardiographic features which are heart rate, heart-rate variability, QRS duration, ST level, QT interval, and corrected QT interval. Results: For athletes 35 years old or younger, preliminary values were [72;91]bpm, [26;47]ms, [85;104]ms, [-0.08;0.08]mm, [326;364]ms and [378;422]ms, respectively. For athletes older than 35 years old, preliminary values were [71;94]bpm, [16;65]ms, [85;100]ms, [-0.11;0.07]mm, [330;368]ms and [394;414]ms, respectively. Conclusion: Availability of preliminary reference values could help identify those athletes who, due to electrocardiographic features out of normal ranges, are more likely to develop cardiac complications that may lead to sport-related sudden cardiac death