Additive-manufactured paper-PMMA hybrid microfluidic chip for simultaneous monitoring of creatinine and pH in artificial urine.

Nowadays, kidney dysfunction is a common health issue due to the modernized lifestyle. Even though medications are commercially available to treat kidney diseases, early diagnosis is crucial and challenging. Clinically, measuring urine creatinine and pH has gained significant interest as a way to diagnose kidney diseases early. In the present work, we attempted to develop a low-cost, robust, accurate and naked-eye colorimetric method to determine both creatinine levels and pH variations in artificial urine samples using a simple 3D-printed hybrid microfluidic device. Creatinine was detected by the incorporation of the traditional Jaffe test onto the hybrid paper-PMMA microfluidic device and pH (4-8) was measured by a simple anthocyanin test. Notably, the tests were established without employing any sophisticated or costly instrument clusters. The developed 3D-printed microfluidic probe showed a limit of detection (LOD) of 0.04 mM for creatinine over a concentration range of 1-10 mM, with a regression coefficient (R2) of 0.995 in laboratory conditions. Interestingly, the experimental data obtained with artificial urine exhibited a wide linear range from 0.1 mM to 5 mM under different pH values ranging from 4 to 8 in the presence of matrices commonly found in urine samples other than proteins, indicating the potential use of this method in pre-clinical analysis. Since the wide linear range of urine creatinine in artificial urine samples falls well below the clinically relevant concentrations in humans (0.07-0.27 mM), the developed lab-on-chip device is further suitable for clinical evaluation with proper ethical clearance. This 3D-printed hybrid microfluidic colorimetry-based creatinine detection and pH indicator platform can be beneficial in the healthcare sector due to the on-site testing capability, cost-effectiveness, ease of use, robustness, and instrument-free approach.

Metal regulation of Mycobacterium tuberculosis SufB intein splicing at the host-pathogen crossroad.

Intein sequences self-excise from precursor proteins to generate functional proteins in various organisms. Thus, regulation of intein splicing at the host-pathogen interface can determine the fate of infection by controlling generation of essential proteins in microbes. For instance, Mycobacterium tuberculosis (Mtu) SufB intein splicing is crucial for the functionality of SUF complex. This multiprotein system is the sole pathway for [Fe-S] cluster biogenesis in mycobacteria during oxidative stress and Fe starvation. Although metal toxicity and metal starvation are components of host immunity, correlation of metal stress to Mtu SufB intein splicing is missing till date. Current study examines the splicing and N-terminal cleavage reactions of Mtu SufB precursor protein in presence of micronutrient metal ions like Zn+2, Cu+2, and Fe+3/+2. A known intein splicing inhibitor Pt+4 was also tested to support its proposed role as an anti-TB agent. Mtu SufB precursor protein exhibited significant attenuation of splicing and N-terminal cleavage reactions across different concentration ranges for Pt+4, Cu+2, Zn+2, while Fe+3 interaction resulted in precursor accumulation. UV-Vis spectroscopy, inductively coupled plasma-optical emission spectroscopy (ICP-OES), Tryptophan fluorescence assay, and dynamic light scattering (DLS) techniques analyzed metal-protein interaction. Mutagenesis experiments and Ellman's assay identified plausible metal co-ordination sites within Mtu SufB protein. Analyzing the metal effect on Mtu SufB splicing may provide elemental information about the fate of mycobacterial infection, and a probable mechanism to attenuate intracellular survival of Mtu. Current research hints at the host regulatory mechanism on SufB splicing in its native environment and a likely target for developing next-generation anti-TB drugs.

Sonophotocatalytic disinfection of Shigella species under visible light irradiation: Insights into its molecular mechanism, antibacterial resistance and biofilm formation.

Microbial contamination of water is one of the major sources of many diseases worldwide. Evolution of antibacterial resistance (ABR) alongside the caveats in most of the water treatment methods causes the severity of the current problem extremely vexing. This calls for an urgent need to develop new treatment methods aiming to reduce the microbial as well as ABR load in the environment. Herein, we successfully developed a visible light assisted sonophotocatalysis (SPC) using Fe/ZnO nanoparticles (NPs) for the disinfection of Shigella dysenteriae. A consortia containing S. dysenteriae and S. flexineri was also completely disinfected using SPC. Growth conditions of S. dysenteriae like growth phases and growth temperaturehad different outcomes on the overall efficacy of SPC. Compared with catalysts such as ZnO and TiO2, Fe/ZnO resulted in better disinfection. Multi-ROS production, mostly containing h+ and O2· radicals, due to the electron displacement in the catalyst and acoustic cavitation was identified as the factors behind bacterial lethality. The ROS produced was found to interfere with the metabolic activities of S. dysenteriae by causing membrane perturbation. We identified DNA damage inside the cells and the subsequent release of intracellular components. The compositional changes in the fatty acid makeup of the cells were altered as a result of SPC and few fatty acid markers indicating the stress posed by SPC were also identified. Loss of ABR in S. dysenteriae was also recorded post SPC treatment. Abatement in the biofilm forming ability of the injured bacterial cells was also recorded, proving the extremity of stress induced by SPC. Hence, the excellent efficacy of SPC in disinfecting bacteria is proposed for tertiary water treatment applications.

Implications of biosensors and nanobiosensors for the eco-friendly detection of public health and agro-based insecticides: A comprehensive review.

Biosensors, in particular nanobiosensors, have brought a paradigm shift in the detection approaches involved in healthcare, agricultural, and industrial sectors. In accordance with the global expansion in the world population, there has been an increase in the application of specific insecticides for maintaining public health and enhancing agriculture, such as organophosphates, organochlorines, pyrethroids, and carbamates. This has led to the contamination of ground water, besides increasing the chances of biomagnification as most of these insecticides are non-biodegradable. Hence, conventional and more advanced approaches are being devised for the routine monitoring of such insecticides in the environment. This review walks through the implications of biosensors and nanobiosensors, which could offer a wide range of benefits for the detection of the insecticides, quantifying their toxicity status, and versatility in application. Unique eco-friendly nanobiosensors such as microcantilevers, carbon nanotubes, 3D printing organic materials and nylon nano-compounds are some advanced tools that are being employed for the detection of specific insecticides under different conditions. Furthermore, in order to implement a smart agriculture system, nanobiosensors could be integrated into mobile apps and GPS systems for controlling farming in remote areas, which would greatly assist the farmer remotely for crop improvement and maintenance. This review discusses about such tools along with more advanced and eco-friendly approaches that are on the verge of development and could offer a promising alternative for analyte detection in different domains.

Template-Free Assembly in Living Bacterial Suspension under an External Electric Field.

Although template-assisted self-assembly methods are very popular in materials and biological systems, they have certain limitations such as lack of tunability and switchable functionality because of the irreversible association of cells and their matrix components. With an aim to achieve more tunability, we have made an attempt to investigate the self-assembly behavior of rod-shaped living bacteria subjected to an external alternating electric field using confocal microscopy. We demonstrate that rod-shaped living bacteria dispersed in a low salinity aqueous medium form different types of reversible freely suspended structures when subjected to an external alternating electric field. At low field strength, an oriented phase is observed where individual bacterium orients with its major axis aligned along the field direction. At intermediate field strength, bacteria align in the form of one-dimensional (1D) chains that lie along the field direction. Further, at high field strength, more bacteria associate with these 1D chains laterally to form a two-dimensional (2D) array. At higher bacterial concentration, these field-induced 2D arrays extend to form three-dimensional columnar structures. These results are discussed in the context of previously reported studies on bacterial self-assembly.