Precision phenylalanine sensing in blood with nanomaterial-enhanced electrodes.

Phenylketonuria (PKU) is a genetic disorder associated with the metabolic dysfunction of the phenylalanine hydroxylase enzyme, which catalyses the conversion of l-phenylalanine (Phe) to l-tyrosine. Elevated levels of phenylalanine disrupt the central nervous system by impairing the myelination process and leading to mental retardation. Currently, commonly used diagnostic methods for PKU include the Guthrie test, liquid chromatography, and tandem mass spectrometry, all of which necessitate sophisticated infrastructure and costly equipment. Conversely, electrochemical detection methods hold promise in clinical diagnosis due to their high accuracy, rapid response time, and user-friendly nature. The choice of electrodes in electrochemical methods significantly influences sensitivity and analytical performance. In this study, we evaluated the performance of various nanomaterial-modified electrodes and compared their responses to the redox reaction of phenylalanine, focusing on detection capabilities in blood samples. Specifically, we examined carbon nanotube-gold nanoparticle modified carbon electrode (C-CNT-GNP), graphene-gold nanoparticle modified carbon electrode (C-GPH-GNP), electrochemically reduced graphene oxide (ERGO) modified carbon electrode (C-ERGO), bare carbon electrode (C-BARE), ERGO modified gold electrode (Au-ERGO), and bare gold electrode (Au-BARE) using amperometric detection. The performance of these electrodes was compared in terms of their limit of detection (LOD), limit of quantification (LOQ), and sensitivity. Among all electrodes, ERGO gold electrode showed the lowest LOD, LOQ, and highest sensitivity. This study highlights the potential of ERGO-modified gold electrodes for enhancing electrocatalytic activity, thus offering promising prospects for further diagnostic applications.

Red cabbage extract-mediated colorimetric sensor for swift, sensitive and economic detection of urease-positive bacteria by naked eye and Smartphone platform.

The bacterial pathogens have caused various serious infectious diseases in the human body, and even some threats to human life by leading to deaths. Enterobacteriaceae species especially urease positive ones, Proteus mirabilis (P. mirabilis) and Klebsiella pneumoniae (K. pneumoniae), show resistance to antibiotics and cause respiratory and urinary tract infections. We have developed natural indicator-incorporated colorimetric urease tests with a naked eye and smartphone readout to rapidly, sensitively and economically detect P. mirabilis and K. pneumoniae. We utilized anthocyanin found as a predominant component in red cabbage (Brassica oleracea) extract as a natural pH indicator instead of toxic and synthetic indicators. As a mechanistic explanation for the detection of P. mirabilis and K. pneumoniae, urease enzymes secreted from the P. mirabilis and K. pneumoniae hydrolyze urea to produce ammonia (NH3), which increases the pH value of the reaction environment and leads to deprotonation from anthocyanins. The changes in the molecular structure and electronic structure of anthocyanins are responsible for revealing many different colors. We demonstrated how some reaction parameters including the concentration of the bacteria (colony-forming unit, CFU), the concentration of anthocyanin in the tests, initial color and pH values (pHs) of the tests influence their detection performance. We further developed a 3D-printed smartphone platform with smartphone based digital image processing software to improve the detection limit and shorten the detection time. We claim that natural indicator-incorporated rapid urease tests providing colorimetric readout evaluated by the human eye and smartphone imaging processing has great potential in practical use and they can be implemented in clinics.

Experimental Investigation of Microfluidic Device for Platelet Activation.

OBJECTIVE: Wound healing is accelerated when Platelet Rich Plasma is activated and growth factors are released. In this study, it was aimed to stimulate platelets without using chemical stimulants. METHOD: Two types of mechanical platelet activation methods have been proposed in this study. The first one is a microfluidic chip developed with the shear-induced platelet activation approach. The second one is a piezo-based ultrasound-assisted device which provides platelet activation by stimulating with an ultrasonic wave (0.55 and 1.1 MHz). Three different microfluidic chip designs were worked out to determine the optimal shear stress characteristics; 8-nodes (2789 µs, 288 shear pulses, and 98.3 dyne/cm2), 40-nodes (2765 µs, 1440 shear pulses, and 95.5 dyne/cm2) and pillar-shaped (1030 µs, 1656 shear pulses, and 48.1 dyne/cm2). RESULTS: The highest platelet activation rate (72.7%) was obtained from the chips with 8-nodes. In the ultrasound-assisted device, 32.4% activation rate was obtained from ultrasound waves with 0.55 MHz frequency and 10 Vp-p amplitude. These activation rates, determined by CD62P (P-Selectin) expression, are significantly higher than spontaneous activation of intact platelets (8.5%). In addition, the gradual increase in activation of stimulated platelets with incubation at room temperature showed that activation continued after stimulation. CONCLUSION: The results showed that these microfluidic devices can be used for platelet activation to enhance the effect of PRP treatment and might reduce adverse immune reactions that may happened due to the use of exogenous activator substances. SIGNIFICANCE: Fast-response, low-cost, easy-to-use and controllable biomedical device have been developed for PRP applications.