Immuno field-effect transistor (ImmunoFET) for detection of salivary cortisol using potentiometric and impedance spectroscopy for monitoring heart failure.

Cortisol, a steroid hormone mostly known as "the stress hormone," plays many essential functions in humans due its involvement in several metabolic pathways. It is well-known that cortisol dysregulation is implied in evolution and progression of several chronic pathologies, including cardiac diseases such as heart failure (HF). However, although several sensors have been proposed to date for the determination of cortisol, none of them has been designed for its determination in saliva in order to monitor HF progression. In this work, a silicon nitride based Immuno field-effect transistor (ImmunoFET) has been proposed to quantify salivary cortisol for HF monitoring. Sensitive biological element was represented by anti-cortisol antibody bound onto the ISFET gate via 11-triethoxysilyl undecanal (TESUD) by vapor-phase method. Potentiometric and electrochemical impedance spectroscopy (EIS) measurements were carried out for preliminary investigations on device responsiveness. Subsequently, a more sensitive detection was obtained using electrochemical EIS. The proposed device has proven to have a linear response (R2 always >0.99), to be sensitive (with a limit of detection, LoD, of 0.005 ± 0.002 ng/mL), selective in case of other HF biomarkers (e.g. N-terminal pro B-type natriuretic peptide (NT-proBNP), tumor necrosis factor-alpha (TNF-α), and interleukin 10 (IL-10)), and accurate in cortisol quantification in saliva sample by performing the standard addition method.

A novel electrochemical strategy for NT-proBNP detection using IMFET for monitoring heart failure by saliva analysis.

Heart failure (HF) is a chronic cardiovascular disease that represents main cause of mortality worldwide, particularly for elderly. N-terminal pro-brain natriuretic peptide (NT-proBNP) was identified as the gold standard biomarker for HF diagnosis and therapy monitoring. Presently, saliva analysis represents an emerging and powerful tool for clinical applications and electrochemical immunosensors have shown their potential in Healthcare applications as selective and reliable systems for detecting clinical biomarkers. This work presents the detection of NT-proBNP in saliva samples by an immunologically modified Field effect Transistor (IMFET). TESUD ((11-triethoxysilyl) undecanal) was used as cross-linker to immobilise anti-NT-proBNP antibody onto the device. Our IMFET that was then tested in different matrices (e.g. phosphate buffered saline (PBS), artificial saliva and human saliva) using electrochemical impedance spectroscopy (EIS), and it resulted selective to NT-proBNP with good sensitivity (detection limit of 0.02 pg/mL) and a wide linear range (0.02-1 pg/mL and 0.5-20 pg/mL). Finally, NT-proBNP concentration in ten saliva samples was determined by performing the standard addition method. An enzyme-linked immunosorbent assay was used for confirming IMFET results, highlighting both IMFET accuracy (analyte recovery of 99 ± 8%) and precision (coefficient of variation always <10%), and supporting the suitability of the device for determining salivary NT-proBNP.

Electrochemical Impedance Spectroscopy Microsensor Based on Molecularly Imprinted Chitosan Film Grafted on a 4-Aminophenylacetic Acid (CMA) Modified Gold Electrode, for the Sensitive Detection of Glyphosate.

A novel electrochemical impedance spectroscopy (EIS) microsensor was implemented for the dosage of traces of glyphosate, in real and synthetic water samples. Molecularly imprinted chitosan was covalently immobilized on the surface of the microelectrode previously modified with 4-aminophenylacetic acid (CMA). The characterization of the resulting microelectrodes was carried out by using cyclic voltammetry measurement (CV), scanning electron microscopy (SEM), and electrochemical impedance spectrometry (EIS). EIS responses of the CS-MIPs/CMA/Au microsensor toward GLY was well-proportional to the concentration in the range from 0.31 × 10-9 to 50 × 10-6 mg/mL indicating a good correlation. The detection limit of GLY was 1 fg/mL (S/N = 3). Moreover, this microsensor showed good reproducibility and repeatability, high selectivity, and can be used for the detection of GLY in river water.

A silicon nitride ISFET based immunosensor for tumor necrosis factor-alpha detection in saliva. A promising tool for heart failure monitoring.

According to the European statistics, approximately 26 million patients worldwide suffer from heart failure (HF), and this number seems to be steadily increasing. Inflammation plays a central role in the development of HF, and the pro-inflammatory cytokine Tumor necrosis factor-α (TNF-α) represents inflammation gold-standard biomarker. Early detection plays a crucial role for the prognosis and treatment of HF. An Ion Sensitive Field Effect Transistor (ISFET) based on silicon nitride transducer and biofunctionalized with anti-TNF-α antibody for label-free detection of salivary TNF-α is proposed. Electrochemical impedance spectroscopy (EIS) was used for TNF-α detection. Our ImmunoFET offered a detection limit of 1 pg mL-1, with an analytical reproducibility expressed by a coefficient of variance (CV) resulted < 10% for the analysis of saliva samples, and an analyte recovery of 94 ± 6%. In addition, it demonstrated high selectivity when compared to other HF biomarkers such as Inteleukin-10, N-terminal pro B-type natriuretic peptide, and Cortisol. Finally, ImmunoFET accuracy in determining the unknown concentration of TNF-α was successfully tested in saliva samples by performing standard addition method. The proposed ImmunoFET showed great promise as a complementary tool for biomedical application for HF monitoring by a non-invasive, rapid and accurate assessment of TNF-α.

Prevalence and severity of groin problems in Spanish football: A prospective study beyond the time-loss approach.

The time-loss definition of injury is commonly adopted in epidemiological groin-injury studies in football, with a significant risk of underestimating the impact of these injuries. This study investigated the extent of groin problems, beyond the time-loss approach, over a full Spanish football season. Players from 17 amateur male teams were followed over 39 consecutive weeks. Groin-injury time loss and self-reported groin pain, irrespective of time loss, were combined to calculate the average weekly prevalence of all groin problems with or without time loss. A subscale measuring hip- and groin-related sporting function from the Copenhagen Hip and Groin Outcome Score questionnaire (HAGOS, Sport/Rec) was registered every 4 weeks. In total, 407 players participated in the study. The average (range) weekly prevalence of all groin problems was 11.7% (7.2%-20.8%); 1.3% with time loss (0.0%-3.2%) and 10.4% without time loss (6.3%-17.6%). Players with groin problems reported lower scores (mean difference) on the HAGOS, Sport/Rec subscale compared with players without (-19.5 [95% CI: -20.7 to -18.4]), while there was no difference between players reporting groin problems with and without time loss (4.0 [95% CI: -1.1 to 9.1]). The traditional time-loss measure only captured 10% of all groin problems. Hip- and groin-related sporting function was not different between players reporting groin problems with or without time loss, suggesting the reason for continuing to play is not only related to the severity of symptoms. These findings question the judicious use of the time-loss approach in overuse conditions, such as groin pain in footballers.