Search Results (3)

Creatinine (Crn) is a clinically relevant biomarker commonly used for the diagnosis and monitoring of kidney disease. In this work, we report the fabrication of reduced-graphene-oxide-based field-effect transistors (rGO FETs) for Crn detection. These devices were functionalized using a layer-by-layer (LbL) assembly, in which polyethyleneimine (PEI) and creatinine deiminase (CD) were alternately deposited. This LbL strategy allows for the effective incorporation of CD without compromising its structural or functional integrity, while also taking advantage of the local pH changes caused by creatinine hydrolysis. It also benefits from the use of a polyelectrolyte that can amplify the enzymatic signal. Furthermore, it enables scalable and efficient fabrication. These transistors also address the challenges of point-of-care implementation in single-use cartridges. It is worth noting that the devices showed a linear relationship between the Dirac-point shift and the logarithm of the creatinine concentration in the 20–500 µM range in diluted simulated urine. The sensor response improved with increasing numbers of PEI/CD bilayers. Furthermore, the functionalized FETs demonstrated rapid detection dynamics and good long-term stability. Present results confirm the potential of these devices as practical biosensors for sample analysis under real-world conditions, making them ideal for implementation in practical settings. Full article

Due to the possible coexistence of Klebsiella pneumoniae (KP) and Candida albicans (CA), strains of KP and CA with biofilm production properties clinically isolated from patients were tested. The production of biofilms from the combined organisms (KP+CA) was higher than the biofilms from each organism alone, as indicated by crystal violet and z-stack immunofluorescence. In parallel, the bacterial abundance in KP + CA was similar to KP, but the fungal abundance was higher than CA (culture method), implying that CA grows better in the presence of KP. Proteomic analysis was performed to compare KP + CA biofilm to KP biofilm alone. With isolated mouse neutrophils (thioglycolate induction), KP + CA biofilms induced less prominent responses than KP biofilms, as determined by (i) neutrophilic supernatant cytokines (ELISA) and (ii) neutrophil extracellular traps (NETs), using immunofluorescent images (neutrophil elastase, myeloperoxidase, and citrullinated histone 3), peptidyl arginine deiminase 4 (PAD4) expression, and cell-free DNA. Likewise, intratracheal KP + CA in C57BL/6 mice induces less severe pneumonia than KP alone, as indicated by organ injury (serum creatinine and alanine transaminase) (colorimetric assays), cytokines (ELISA), bronchoalveolar lavage fluid parameters (bacterial culture and neutrophil abundances using a hemocytometer), histology score (H&E stains), and NETs (immunofluorescence on the lung tissue). In conclusion, the biofilm biomass of KP + CA was mostly produced from CA with less potent neutrophil activation and less severe pneumonia than KP alone. Hence, fungi in the respiratory tract might benefit the host in some situations, despite the well-known adverse effects of fungi. Full article

Urine analysis is widely used in clinical practice to indicate human heathy status and is important for diagnosing chronic kidney disease (CKD). Ammonium ions (NH₄⁺), urea, and creatinine metabolites are main clinical indicators in urine analysis of CKD patients. In this paper, NH₄⁺ selective electrodes were prepared using electropolymerized polyaniline-polystyrene sulfonate (PANI: PSS), and urea- and creatinine-sensing electrodes were prepared by modifying urease and creatinine deiminase, respectively. First, PANI: PSS was modified on the surface of an AuNPs-modified screen-printed electrode, as a NH₄⁺-sensitive film. The experimental results showed that the detection range of the NH₄⁺ selective electrode was 0.5~40 mM, and the sensitivity reached 192.6 mA M⁻¹ cm⁻² with good selectivity, consistency, and stability. Based on the NH₄⁺-sensitive film, urease and creatinine deaminase were modified by enzyme immobilization technology to achieve urea and creatinine detection, respectively. Finally, we further integrated NH₄⁺, urea, and creatinine electrodes into a paper-based device and tested real human urine samples. In summary, this multi-parameter urine testing device offers the potential for point-of-care testing of urine and benefits the efficient chronic kidney disease management. Full article