Detecting Dementia by Saliva Analysis: A Fingerprinting Unobtrusive Method Based on a Fast and Cheap Sensor System.

BACKGROUND: Recent studies have shown that oxidative stress plays a relevant role in Alzheimer's disease (AD), and in the pathogenesis of vascular dementia (VaD). New diagnostic methods look for biological samples with non-invasive sampling methods. Among these, saliva shows an increase in oxidative stress products, thus a corresponding reduction in antioxidant products were found in dementia cases compared to healthy controls. Compounds identified in saliva include some hydrocarbons whose production has been related to the presence of reactive oxygen species. OBJECTIVE: The hypothesis is that the voltammetric analysis performed on saliva could be a useful test for diagnosing dementia, potentially discriminating between AD and VaD. METHODS: A single-center observational study was conducted on patients referred to the dementia clinic in the Neurology area and healthy controls recruited in the Orthopedics area of the Campus Bio-Medico Hospital in Rome. The study was aimed at evaluating the discriminative properties of salivary voltammetric analysis between healthy subjects and patients with dementia and, as a secondary outcome, between AD and VaD. A total of 69 subjects were enrolled, including 29 healthy controls, 20 patients with AD, and 20 patients with VaD. The degree of cognitive impairment was classified on the basis of the Mini-Mental State Examination score. RESULTS: The results obtained are promising, with an accuracy of 79.7%, a sensitivity of 82.5%, and a specificity of 75.8%, in the discrimination of dementia versus controls. CONCLUSIONS: The methods tested demonstrate to be relevant in the discrimination between dementia and controls. A confirmatory study is already running.

Microsystem Nodes for Soil Monitoring via an Energy Mapping Network: A Proof-of-Concept Preliminary Study.

The need for accurate information and the availability of novel tool and technological advances in agriculture have given rise to innovative autonomous systems. The aim is to monitor key parameters for optimal water and fertilizer management. A key issue in precision agriculture is the in situ monitoring of soil macronutrients. Here, a proof-of-concept study was conducted that tested two types of sensors capable of capturing both the electrochemical response of the soil and the electrical potential generated by the interaction between the soil and plants. These two sensors can be used to monitor large areas using a network approach, due to their small size and low power consumption. The voltammetric sensor (BIONOTE-L) proved to be able to characterize different soil samples. It was able, indeed, to provide a reproducible voltammetric fingerprint specific for each soil type, and to monitor the concentration of CaCl2 and NaCl in the soil. BIONOTE-L can be coupled to a device capable of capturing the energy produced by interactions between plants and soil. As a consequence, the functionality of the microsystem node when applied in a large-area monitoring network can be extended. Additional calibrations will be performed to fully characterize the instrument node, to implement the network, and to specialize it for a particular application in the field.

Micromachined Tools Using Acoustic Wave Triggering for the Interaction with the Growth of Plant Biological Systems.

A plant biological system is exposed to external influences. In general, each plant has its characteristics and needs with specific interaction mechanisms adapted to its survival. Interactions between systems can be examined and modeled as energy exchanges of mechanical, chemical or electrical variables. Thus, each specific interaction can be examined by triggering the system via a specific stimulus. The objective of this work was to study a specific stimulus (mechanical stimulation) as a driver of plants and their interaction with the environment. In particular, the experimental design concerns the setting up and testing of an automatic source of mechanical stimuli at different wavelengths, generated by an electromechanical transducer, to induce a micro-interaction in plants (or in parts of them) that produces a specific behavior (hypothesis) of plants. Four different experimental setups were developed for this work, each pursuing the same objective: the analysis of the germination process induced by stimulation by sound waves in the audible range. It can be said that the introduction of sound waves as a stimulant or a brake for the growth of plants can offer significant advantages when used on a large scale in the primary sector, since these effects can be used instead of polluting chemical solutions.

A Multi-Sensor System for Sea Water Iodide Monitoring and Seafood Quality Assurance: Proof-of-Concept Study.

Iodine is a trace chemical element fundamental for a healthy human organism. Iodine deficiency affects about 2 billion people worldwide causing from mild to severe neurological impairment, especially in children. Nevertheless, an adequate nutritional intake is considered the best approach to prevent such disorders. Iodine is present in seawater and seafood, and its common forms in the diet are iodide and iodate; most iodide in seawater is caused by the biological reduction of the thermodynamically stable iodate species. On this basis, a multisensor instrument which is able to perform a multidimensional assessment, evaluating iodide content in seawater and seafood (via an electrochemical sensor) and discriminating when the seafood is fresh or defrosted quality (via a Quartz Micro balance (QMB)-based volatile and gas sensor), is strategic for seafood quality assurance. Moreover, an electronic interface has been opportunely designed and simulated for a low-power portable release of the device, which should be able to identify seafood over or under an iodide threshold previously selected. The electrochemical sensor has been successfully calibrated in the range 10-640 µg/L, obtaining a root mean square error in cross validation (RMSECV) of only 1.6 µg/L. Fresh and defrosted samples of cod, sea bream and blue whiting fish have been correctly discriminated. This proof-of-concept work has demonstrated the feasibility of the proposed application which must be replicated in a real scenario.

Development and Test of a Portable ECG Device with Dry Capacitive Electrodes and Driven Right Leg Circuit.

The use of wearable sensors for health monitoring is rapidly growing. Over the past decade, wearable technology has gained much attention from the tech industry for commercial reasons and the interest of researchers and clinicians for reasons related to its potential benefit on patients' health. Wearable devices use advanced and specialized sensors able to monitor not only activity parameters, such as heart rate or step count, but also physiological parameters, such as heart electrical activity or blood pressure. Electrocardiogram (ECG) monitoring is becoming one of the most attractive health-related features of modern smartwatches, and, because cardiovascular disease (CVD) is one of the leading causes of death globally, the use of a smartwatch to monitor patients could greatly impact the disease outcomes on health care systems. Commercial wearable devices are able to record just single-lead ECG using a couple of metallic contact dry electrodes. This kind of measurement can be used only for arrhythmia diagnosis. For the diagnosis of other cardiac disorders, additional ECG leads are required. In this study, we characterized an electronic interface to be used with multiple contactless capacitive electrodes in order to develop a wearable ECG device able to perform several lead measurements. We verified the ability of the electronic interface to amplify differential biopotentials and to reject common-mode signals produced by electromagnetic interference (EMI). We developed a portable device based on the studied electronic interface that represents a prototype system for further developments. We evaluated the performances of the developed device. The signal-to-noise ratio of the output signal is favorable, and all the features needed for a clinical evaluation (P waves, QRS complexes and T waves) are clearly readable.

CO2 and O2 Detection by Electric Field Sensors.

In this work an array of chemical sensors for gas detection has been developed, starting with a commercial sensor platform developed by Microchip (GestIC), which is normally used to detect, trace, and classify hand movements in space. The system is based on electric field changes, and in this work, it has been used as mechanism revealing the adsorption of chemical species CO2 and O2. The system is composed of five electrodes, and their responses were obtained by interfacing the sensors with an acquisition board based on an ATMEGA 328 microprocessor (Atmel MEGA AVR microcontroller). A dedicated measurement chamber was designed and prototyped in acrylonitrile butadiene styrene (ABS) using an Ultimaker3 3D printer. The measurement cell size is 120 × 85 mm. Anthocyanins (red rose) were used as a sensing material in order to functionalize the sensor surface. The sensor was calibrated using different concentrations of oxygen and carbon dioxide, ranging from 5% to 25%, mixed with water vapor in the range from 50% to 90%. The sensor exhibits good repeatability for CO2 concentrations. To better understand the sensor response characteristics, sensitivity and resolution were calculated from the response curves at different working points. The sensitivity is in the order of magnitude of tens to hundreds of µV/% for CO2, and of µV/% in the case of O2. The resolution is in the range of 10-1%-10-3% for CO2, and it is around 10-1% for O2. The system could be specialized for different fields, for environmental, medical, and food applications.

A Sensor Platform for Athletes' Training Supervision: A Proof of Concept Study.

One of the basic needs of professional athletes is the real-time and non-invasive monitoring of their activities. The use of these kind of data is necessary to develop strategies for specific tailored training in order to improve performances. The sensor system presented in this work has the aim to adopt a novel approach for the monitoring of physiological parameters, and athletes' performances, during their training. The anaerobic threshold is herein identified with the monitoring of the lactate concentration and the respiratory parameters. The data collected by the sensor are used to build a model using a supervised method (based on the partial least squares method, PLS) to predict the values of the parameters of interest. The sensor is able to measure the lactate concentration from a sample of saliva and it can estimate a respiratory parameter, such as maximal oxygen consumption, maximal carbon dioxide production and respiratory rate from a sample of exhaled breath. The main advantages of the device are the low power; the wireless communication; and the non-invasive sampling method, which allow its use in a real context of sport practice.

Exhaled breath analysis in hepatology: State-of-the-art and perspectives.

Liver disease is characterized by breath exhalation of peculiar volatile organic compounds (VOCs). Thanks to the availability of sensitive technologies for breath analysis, this empiric approach has recently gained increasing attention in the context of hepatology, following the good results obtained in other fields of medicine. After the first studies that led to the identification of selected VOCs for pathophysiological purposes, subsequent research has progressively turned towards the comprehensive assessment of exhaled breath for potential clinical application. Specific VOC patterns were found to discriminate subjects with liver cirrhosis, to rate disease severity, and, eventually, to forecast adverse clinical outcomes even beyond existing scores. Preliminary results suggest that breath analysis could be useful also for detecting and staging hepatic encephalopathy and for predicting steatohepatitis in patients with nonalcoholic fatty liver disease. However, clinical translation is still hampered by a number of methodological limitations, including the lack of standardization and the consequent poor comparability between studies and the absence of external validation of obtained results. Given the low-cost and easy execution at bedside of the new technologies (e-nose), larger and well-structured studies are expected in order to provide the adequate level of evidence to support VOC analysis in clinical practice.

Repeatability of exhaled breath fingerprint collected by a modern sampling system in asthmatic and healthy children.

E-noses provide potential non-invasive metabolic biomarkers for diagnosing and monitoring pulmonary diseases. The primary aim of the present study was to assess the within-day and between-day repeatability of a modern breath sampling system (Pneumopipe® plus an array of e-nose sensors) in asthmatic and healthy children. The secondary aim was to compare the repeatability of the breath sampling system, spirometry and exhaled nitric oxide (eNO). Fifteen children (age 6-11 years) with asthma and thirty healthy children matched by age and gender (1:2 allocation) were recruited; of them, three healthy children did not complete the study. All measurements were collected twice during the baseline visit, 30 min apart, and once during the final visit, after 7 d. Repeatability was assessed through the intra-cluster correlation coefficient (ICC), and a significance test was performed to detect an at least 'fair' repeatability (ICC > 0.2). In asthmatic children, the within-day (0-30 min) ICCs for e-nose sensors (8 sensors × 4 desorption temperatures) ranged from 0.24 to 0.84 (median 0.57, IQR 0.47-0.71), while the between-day (0-7 d) ICCs ranged from 0.25 to 0.83 (median 0.66, IQR 0.55-0.72). In healthy children, the within-day ICCs for e-nose sensors ranged from 0.29 to 0.85 (median 0.58, IQR 0.49-0.63), while the between-day ICCs ranged from 0.33 to 0.82 (median 0.55, IQR 0.49-0.63). In both groups, most of the within-day and between-day ICCs for e-nose sensors were statistically significant. Moreover, the within-day and between-day ICCs for all spirometry parameters and eNO were significant and similar to those of the most reliable sensors. The modern breath sampling system showed more than acceptable within-day and between-day repeatability, in both asthmatic and healthy children. The present study was registered on the central registration system ClinicalTrials.gov (ID: NCT03025061).

Advances in the Electronics for Cyclic Voltammetry: the Case of Gas Detection by Using Microfabricated Electrodes.

This paper presents an advanced voltammetric system to be used as electronic tongue for liquid and gas analysis. It has been designed to be more flexible and accurate with respect to other existing and similar systems. It features improved electronics and additional operative conditions. Among others these include the possibility to optically excite the solution and to treat the output signal by a differentiation process in order to better evidence the existence of small details in the response curve. Finally by the same type of tongue preliminary results are shown dealing with O2 and CO2 concentration measurements in appropriate solutions.

Monitoring of polycyclic aromatic hydrocarbons in bees (Apis mellifera) and honey in urban areas and wildlife reserves.

The honeybee is a good biological indicator that quickly reflects chemical impairment of the environment by its high mortality and the presence of pollutants in its body or in beehive products. In this work the honeybee (Apis mellifera) and honey were used to detect the presence of polycyclic aromatic hydrocarbons (PAHs) in several areas with different degrees of environmental pollution. All sampling sites showed the presence of PAHs. Benzo(a)pyrene was never detected. Fluorene, phenanthrene, anthracene, fluoranthene, benz(a)anthracene, benzo(b)fluoranthene, and benzo(k)fluoranthene were the PAHs detected in bees, whereas the honey contained only phenanthrene, anthracene, and chrysene. Phenanthrene showed the highest mean values in honeybees and honey. Independent from the season and location the pattern of PAHs in honeybees and honey was dominated by the presence of the lowest molecular weight PAHs. Furthermore, the mean PAH concentrations in honey samples were lower than those reported in honeybees, and no positive correlation was found between the compounds detected in bees and those in honey.

Use of quinoline alkaloids as markers of the floral origin of chestnut honey.

To identify potential floral markers of chestnut honey, the phytochemicals present in chestnut floral nectar collected by bees were analyzed. Two nitrogen-containing compounds were detected, isolated, and identified as 4-hydroxyquinaldic acid (kynurenic acid) and 4-quinolone-2-carboxylic acid by (1)H NMR and (13)C NMR. In addition, chestnut nectar contained the monoterpene 4-(1-hydroxy-1-methylethyl)cyclohexa-1,3-diene-1-carboxylic acid, its gentiobioside ester, and the flavonol quercetin 3-pentosylhexoside. These nectar markers were found in different chestnut unifloral honey samples, although the flavonol was not detected in all samples analyzed. The terpenoid derivatives had previously been found in linden and tilia honeys. These results show that quinoline alkaloids are potentially good markers of chestnut honey, as they were not detected in any other unifloral honey analyzed so far. They are present at concentrations ranging from 34 to 65 mg/100 g of honey in the samples analyzed. In addition, the terpenoid and flavonoid derivatives present in nectar, although not exclusively characteristic of this floral origin, are good complementary markers for the determination of the floral origin of chestnut honey.

Nectar Flavonol rhamnosides are floral markers of acacia (Robinia pseudacacia) honey.

With the objective of finding floral markers for the determination of the botanical origin of acacia (robinia) honey, the phytochemicals present in nectar collected from Robinia pseudacacia flowers were analyzed by high-performance liquid chromatography-tandem mass spectrometry. Eight flavonoid glycosides were detected and characterized as kaempferol combinations with rhamnose and hexose. Acacia honey produced in the same location where the nectar was collected contained nectar-derived kaempferol rhamnosides. This is the first time that flavonoid glycosides have been found as honey constituents. Differences in the stability of nectar flavonoids during honey elaboration and ripening in the hive were shown to be due to hydrolytic enzymatic activity and to oxidation probably related to hydrogen peroxide (glucose-oxidase) activity. Acacia honeys contained propolis-derived flavonoid aglycones (468-4348 microg/100 g) and hydroxycinnamic acid derivatives (281-3249 microg/100 g). In addition, nectar-derived kaempferol glycosides were detected in all of the acacia honey samples analyzed (100-800 microg/100 g). These flavonoids were not detected in any of the different honey samples analyzed previously from different floral origins other than acacia. Finding flavonoid glycosides in honey related to floral origin is particularly relevant as it considerably enlarges the number of possible suitable markers to be used for the determination of the floral origin of honeys.

Classification of Italian honeys by 2D HR-NMR.

The importance of honey has been recently increased because of its nutrient and therapeutic effects, but the adulteration of honey in terms of botanical origin has increased, too. The floral origin of honeys is usually determined using melisso-palynological analysis and organoleptic characteristics, but the application of these techniques requires some expertise. A number of papers have confirmed the possibility of characterizing honey samples by selected chemical parameters. In this study high-resolution nuclear magnetic resonance (HR-NMR) and multivariate statistical analysis methods were used to identify and classify honeys of five different floral sources. The 71 honey samples (robinia, chestnut, citrus, eucalyptus, polyfloral) were analyzed by HR-NMR using both 1H NMR and heteronuclear multiple bond correlation spectroscopy (HMBC). Spectral data were analyzed by application of unsupervised and supervised pattern recognition and multivariate statistical techniques such as principal component analysis (PCA) and general discriminant analysis (GDA). The use of 1H-(13)C HMBC coupled with appropriate statistical analysis seems to be an efficient technique for the classification of honeys.

Determination of Thiamethoxam residues in honeybees by high performance liquid chromatography with an electrochemical detector and post-column photochemical reactor.

A method for the determination of Thiamethoxam in bee samples was set up by means of high performance liquid chromatography with an electrochemical detector and post-column photochemical reactor (HPLC-h nu-ED). Analytical method was based on a rapid sample extraction procedure with acetone, followed by chromatographic separation into a C18-RP column isocratically operated by 60 mM phosphate buffer/acetonitrile (75/25) mobile phase at pH 2.7. A photochemical reactor was used as a tool to verify and eventually quantify the presence of Thiamethoxam in the samples by distinguishing it from interference contribution. Detection was performed with a potential of 880 mV after a photoactivation with a 254 nm light. The least detectable dose was 0.002 mg kg(-1). Recovery rates ranged between 59.88 and 71.62%.

Towards royal jelly proteome.

The recent availability of the honey-bee Apis mellifera genome and trascriptome of both the female castes, has stimulated new efforts in investigating the protein composition of royal jelly (RJ), its role in caste differentiation and its quality and typicality by a proteomic approach. This study is aimed both to separate and identify proteins of royal jelly and to detect some of them in honey-bee pollen-bread by using two-dimensional gel electrophoresis, mass spectrometry and by de novo sequencing. All the identified proteins belonged to the Apis mellifera genome. Apalbumin 1 was also confirmed to be present in honey-bee pollen-bread where the presence of apalbumin 2 was also found. In addition several fragments of apalbumin 1 and apalbumin 3 were also found in RJ. These could be the result of protease activity other than that of serine-protease. This study is a contribution to the description of royal jelly proteome.

Validated methods for the quantification of biologically active constituents of poplar-type propolis.

The validation of rapid, low-cost spectrophotometric procedures for the quantification of the three main groups of bioactive substances (flavones and flavonols, flavanones and dihydroflavonols, and total phenolics) in poplar-type propolis has been performed. A spectrophotometric assay based on the formation of an aluminium chloride complex was applied for the quantification of total flavones and flavonols using galangin as standard. Because of the high amount of flavanones and dihydroflavonols in "poplar type" propolis, the introduction of a distinct procedure for their quantification was considered of special significance and the DAB9 colorimetric method was applied for the purpose. Total phenolic content was measured by the Folin-Ciocalteu procedure using a mixture of pinocembrin and galangin as a reference. The procedures were validated using a model mixture of compounds representing the poplar-type propolis composition as found in previous studies. The accuracy (recovery) varied in the range 84-109%, and the relative standard deviation was 0.5-6.2%. The developed spectrophotometric procedures were applied to six poplar type propolis samples. The results were verified independently by a HPLC procedure. The two sets of results agreed satisfactory, as proven by Student's t-test.

Chemical composition of European propolis: expected and unexpected results.

Ten propolis samples from Bulgaria, Italy and Switzerland were analyzed by GC-MS. As expected, most samples displayed the typical chemical pattern of "poplar" propolis: they contained pinocembrin, pinobanksin and its 3-O-acetate, chrysin, galangin, prenyl esters of caffeic and ferulic acids. Two samples differed significantly: one from the Graubünden Alpine region, Switzerland, rich in phenolic glycerides, and one from Sicily which contained only a limited number of phenolics and was rich in diterpenic acids.