Dopamine-Assisted Synthesis of Carbon-Coated Silica for PCR Enhancement

Polymerase chain reaction (PCR) has become one of the most popular methods to identify genomic information on cells and tissues as well as to solve crimes and check genetic diseases. Recently, the nanomaterials including nanocomposite and nanoparticles have been considered as a next generation of solution to improve both quality and productivity of PCR. Herein, taking into these demands, carbon-coated silica was synthesized using silica particles via polymerization of biocompatible dopamine (PD) to form polydopamine (PDA) film and carbonization of PDA into graphitic structures. For further investigation of the effects of as-prepared silica, PDA-coated silica (PDA silica), and carbonized PDA silica (C-PDA silica), two different types of genes were adopted to investigate the influences of them in the PCR. Furthermore, the strong interaction between the nanocomposites and PCR reagents including polymerase and primers enables regulation of the PCR performance. The effectiveness of the nanocomposites was also confirmed through adopting the conventional PCR and real-time PCR with two different types of DNA as realistic models and different kinds of analytical methods. These findings could provide helpful insight for the potential application in biosensors and biomedical diagnosis

Portable Agrichemical Detection System for Enhancing the Safety of Agricultural Products Using Aggregation of Gold Nanoparticles

Organophosphorus (OP) and triazole chemicals have been commonly used as insecticides and fungicides to protect agricultural foods from harmful insects and fungi. However, these agrichemicals sometimes remain after distribution and can cause serious health and environmental issues. Therefore, it is essential to detect OPs and triazole chemicals in agricultural products. Nowadays, many detection techniques for OPs and triazole chemicals are expensive and time-consuming and require highly trained technicians. Thus, particularly rapid, simple, and sensitive detection methods are in demand for on-site screening of agrichemicals. Gold nanoparticles (AuNPs) have been utilized for applications in analytical assays and real-time monitoring in the biosensor field because of their biocompatibility and outstanding size-dependent optical properties. In this study, we used AuNPs as a detection probe, which have a size of 17 nm in diameter, a red color, and the absorbance peak at 520 nm. When imidazole was added to AuNPs mixed with the agrichemicals, the AuNPs aggregated and their colors changed to purple, causing the appearance of a new peak at 660–670 nm, which could be measured within approximately 20 s. Moreover, we developed a novel device for multiple agrichemical detections using an AuNP-aggregation-based spectrometric detection system. This portable device is light, simple, fast, and highly sensitive as well as selective. With this system, agrichemical residues can be easily detected on the spot at a low cost and in a short reaction time

Cell-Based Method Utilizing Fluorescent <i>Escherichia coli</i> Auxotrophs for Quantification of Multiple Amino Acids

A cell-based assay system for simultaneous quantification of the three amino acids, phenylalanine (Phe), methionine (Met), and leucine (Leu) in a single biological sample, was developed and applied in the multiplex diagnosis of three key metabolic diseases of newborn babies. The assay utilizes three Escherichia coli auxotrophs, which grow only in the presence of the corresponding target amino acids and which contain three different fluorescent reporter plasmids that produce distinguishable fluorescence signals (red, green, and cyan) in concert with cell growth. To mixtures of the three auxotrophs, immobilized on agarose gels arrayed on a well plate, is added a test sample. Following incubation, the concentrations of the three amino acids in the sample are simultaneously determined by measuring the intensities of three fluorescence signals that correspond to the reporter plasmids. The clinical utility of this assay system was demonstrated by employing it to identify metabolic diseases of newborn babies through the quantification of Phe, Met, and Leu in clinically derived dried blood spot specimens. The general strategy developed in this effort should be applicable to the design of new assay systems for the quantification of multiple amino acids derived from complex biological samples and, as such, to expand the utilization of cell-based analytical systems that replace conventional, yet laborious methods currently in use

Enhanced Pseudocapacitance of Ionic Liquid/Cobalt Hydroxide Nanohybrids

Development of nanostructured materials with enhanced redox reaction capabilities is important for achieving high energy and power densities in energy storage systems. Here, we demonstrate that the nanohybridization of ionic liquids (ILs, 1-butyl-3-methylimidazolium tetrafluoroborate) and cobalt hydroxide (Co(OH)₂) through ionothermal synthesis leads to a rapid and reversible redox reaction. The as-synthesized IL-Co(OH)₂ has a favorable, tailored morphology with a large surface area of 400.4 m²/g and a mesopore size of 4.8 nm. In particular, the IL-Co(OH)₂-based electrode exhibits improvement in electrochemical characteristics compared with bare Co(OH)₂, showing a high specific capacitance of 859 F/g at 1 A/g, high-rate capability (∼95% retention at 30 A/g), and excellent cycling performance (∼96% retention over 1000 cycles). AC impedance analysis demonstrates that the introduction of ILs on Co(OH)₂ facilitates ion transport and charge transfer: IL-Co(OH)₂ shows a higher ion diffusion coefficient (1.06 × 10^–11 cm²/s) and lower charge transfer resistance (1.53 Ω) than those of bare Co(OH)₂ (2.55 × 10^–12 cm²/s and 2.59 Ω). Our density functional theory (DFT) calculations reveal that the IL molecules, consisting of anion and cation groups, enable easier hydrogen desorption/adsorption process, that is, a more favorable redox reaction on the Co(OH)₂ surface

Wrinkled Surface-Mediated Antibacterial Activity of Graphene Oxide Nanosheets

Surface wrinkles are commonly observed in large-scale of graphene films. As a new feature, the wrinkled surface of graphene films may directly affect bacterial viability by means of various interactions of bacterial cells with graphene sheets. In the present study, we introduce a wrinkled surface geometry of graphene oxide (GO) thin films for antibacterial application. Highly wrinkled GO films were formed by vacuum filtration of a GO suspension through a prestrained filter. Several types of wrinkled GO surfaces were obtained with different roughness grades determined by root-mean-square values. Antibacterial activity of the fabricated GO films toward three different bacterial species, <i>Escherichia coli</i>, <i>Mycobacterium smegmatis</i>, and <i>Staphylococcus aureus</i>, was evaluated in relation to surface roughness. Because of their nanoscopically corrugated nature, the wrinkled GO films exhibited excellent antibacterial properties. On the basis of our detailed observations, we propose a novel concept of the surrounded contact-based mechanism for antimicrobial activity of wrinkled GO films. It postulates formation of a mechanically robust GO surface “trap” that prompts interaction of bacteria with the diameter-matched GO sink, which results in substantial damages to the bacterial cell membrane. We believe that our approach uncovered a novel use of a promising two-dimensional material for highly effective antibacterial treatment

<i>In Vitro</i> Biosynthesis of Metal Nanoparticles in Microdroplets

We report the use of a hydrogel polymer, recombinant <i>Escherichia coli</i> cell extracts, and a microdroplet-based microfluidic device to fabricate artificial cellular bioreactors which act as reactors to synthesize diverse metal nanoparticles (NPs). The combination of cell extracts, microdroplet-based microfluidic device, and hydrogel was able to produce a mass amount of artificial cellular bioreactors with uniform size and shape. For the first time, we report the alternating generation of microdroplets through one orifice for the fabrication of the artificial cellular reactors using the cell extract as inner cellular components and hydrogel as an artificial cellular membrane. Notably, the hydrogels were able to protect the encapsulated cell extracts from the surrounding environment and maintain the functionality of cellular component for the further cellular bioreactor applications. Furthermore, the successful applications of the fabricated artificial cellular bioreactors to synthesize various NPs including quantum dots, iron, and gold was demonstrated. By employing this microfluidic technique, the artificial cellular bioreactors could be applicable for the synthesis of diverse metal NPs through simple dipping of the reactors to the metal precursor solutions. Thus, the different size of NPs can be synthesized through controlling the concentration of metal precursors. This artificial cellular bioreactors offer promising abilities to biofriendly ways to synthesis diverse NPs and can be applicable in chemical, biomedical, and bioengineering applications

<i>In Vitro</i> Biosynthesis of Metal Nanoparticles in Microdroplets

We report the use of a hydrogel polymer, recombinant <i>Escherichia coli</i> cell extracts, and a microdroplet-based microfluidic device to fabricate artificial cellular bioreactors which act as reactors to synthesize diverse metal nanoparticles (NPs). The combination of cell extracts, microdroplet-based microfluidic device, and hydrogel was able to produce a mass amount of artificial cellular bioreactors with uniform size and shape. For the first time, we report the alternating generation of microdroplets through one orifice for the fabrication of the artificial cellular reactors using the cell extract as inner cellular components and hydrogel as an artificial cellular membrane. Notably, the hydrogels were able to protect the encapsulated cell extracts from the surrounding environment and maintain the functionality of cellular component for the further cellular bioreactor applications. Furthermore, the successful applications of the fabricated artificial cellular bioreactors to synthesize various NPs including quantum dots, iron, and gold was demonstrated. By employing this microfluidic technique, the artificial cellular bioreactors could be applicable for the synthesis of diverse metal NPs through simple dipping of the reactors to the metal precursor solutions. Thus, the different size of NPs can be synthesized through controlling the concentration of metal precursors. This artificial cellular bioreactors offer promising abilities to biofriendly ways to synthesis diverse NPs and can be applicable in chemical, biomedical, and bioengineering applications

Plastic-Chip-Based Magnetophoretic Immunoassay for Point-of-Care Diagnosis of Tuberculosis

Tuberculosis (TB) remains a relevant infectious disease in the 21st century, and its extermination is still far from being attained. Due to the extreme infectivity of incipient TB patients, a rapid sensing system for proficient point-of-care (POC) diagnostics is required. In our study, a plastic-chip-based magnetophoretic immunoassay (pcMPI) is introduced using magnetic and gold nanoparticles (NPs) modified with <i>Mycobacterium tuberculosis</i> (MTB) antibodies. This pcMPI offers an ultrasensitive limit of detection (LOD) of 1.8 pg·ml^–1 for the detection of CFP-10, an MTB-secreted antigen, as a potential TB biomarker with high specificity. In addition, by combining the plastic chip with an automated spectrophotometer setup, advantages include ease of operation, rapid time to results (1 h), and cost-effectiveness. Furthermore, the pcMPI results using clinical sputum culture filtrate samples are competitively compared with and integrated with clinical data collected from conventional tools such as the acid-fast bacilli (AFB) test, mycobacteria growth indicator tube (MGIT), polymerase chain reaction (PCR), and physiological results. CFP-10 concentrations were consistently higher in patients diagnosed with MTB infection than those seen in patients infected with nontuberculosis mycobacteria (NTM) (<i>P</i> < 0.05), and this novel test can distinguish MTB and NTM while MGIT cannot. All these results indicate that this pcMPI has the potential to become a new commercial TB diagnostic POC platform in view of its sensitivity, portability, and affordability

Plastic-Chip-Based Magnetophoretic Immunoassay for Point-of-Care Diagnosis of Tuberculosis

Tuberculosis (TB) remains a relevant infectious disease in the 21st century, and its extermination is still far from being attained. Due to the extreme infectivity of incipient TB patients, a rapid sensing system for proficient point-of-care (POC) diagnostics is required. In our study, a plastic-chip-based magnetophoretic immunoassay (pcMPI) is introduced using magnetic and gold nanoparticles (NPs) modified with <i>Mycobacterium tuberculosis</i> (MTB) antibodies. This pcMPI offers an ultrasensitive limit of detection (LOD) of 1.8 pg·ml^–1 for the detection of CFP-10, an MTB-secreted antigen, as a potential TB biomarker with high specificity. In addition, by combining the plastic chip with an automated spectrophotometer setup, advantages include ease of operation, rapid time to results (1 h), and cost-effectiveness. Furthermore, the pcMPI results using clinical sputum culture filtrate samples are competitively compared with and integrated with clinical data collected from conventional tools such as the acid-fast bacilli (AFB) test, mycobacteria growth indicator tube (MGIT), polymerase chain reaction (PCR), and physiological results. CFP-10 concentrations were consistently higher in patients diagnosed with MTB infection than those seen in patients infected with nontuberculosis mycobacteria (NTM) (<i>P</i> < 0.05), and this novel test can distinguish MTB and NTM while MGIT cannot. All these results indicate that this pcMPI has the potential to become a new commercial TB diagnostic POC platform in view of its sensitivity, portability, and affordability