Carbon electrode obtained via pyrolysis of plasma-deposited parylene-C for electrochemical immunoassays.

In this study, parylene-C films from plasma deposition as well as thermal deposition were pyrolyzed to prepare a carbon electrode for application in electrochemical immunoassays. Plasma deposition could prepare parylene-C in a faster deposition rate and more precise control over the thickness in comparison with the conventional thermal deposition. To analyze the influence of the deposition method, the crystal and electronic structures of the pyrolyzed parylene-C films obtained via both deposition methods were compared using Fourier transform infrared spectroscopy, X-ray photoelectron spectroscopy, and Raman spectroscopy. For application as a carbon electrode in immunoassays, the electrochemical properties of the pyrolyzed carbon films from two both deposition methods were analyzed, including the double layer capacitance (2.10 µF cm-2 for plasma deposition and 2.20 µF cm-2 for thermal deposition), the apparent electron transfer rate (approximately 1.1 × 10-3 cm s-1 for both methods), and the electrochemical window (approximately -1.0 ∼ 2.1 V for both methods). Finally, the applicability of the pyrolyzed carbon electrode from parylene-C was demonstrated for the diagnosis of human hepatitis-C using various amperometric methods, such as cyclic voltammetry, chronoamperometry, square-wave voltammetry and differential pulse voltammetry.

Biodegradation of Polystyrene by Pseudomonas sp. Isolated from the Gut of Superworms (Larvae of Zophobas atratus).

Recently, various attempts have been made to solve plastic waste problems, such as development of biodegradation without producing pollution. Polystyrene (PS) is the fifth most used plastic in many industries; therefore, degrading PS becomes a critical global issue. Here, we reported Pseudomonas aeruginosa strain DSM 50071, initially isolated from the gut of the superworms, Zophobas atratus, and the PS degradation by Pseudomonas sp. DSM 50071. We examined PS degradation using electronic microscopy and measured changes in atomic composition and contact angles with water droplets on the PS surface that represents a chemical change from hydrophobicity to hydrophilicity. We have further examined chemical structural changes using X-ray photoelectron spectroscopy, Fourier-transform-infrared spectroscopy, and nuclear magnetic resonance (NMR) to confirm the formation of carbonyl groups (C═O) in the oxidation pathway during PS biodegradation. In reverse transcription quantitative polymerase chain reaction analysis, the gene expression level of serine hydrolase (SH) in Pseudomonas sp. DSM 50071 was highly increased during PS degradation, and the enzyme-mediated biodegradation of PS was further confirmed by the SH inhibitor treatment test. Thus, the significance of these findings goes beyond the discovery of a novel function of Pseudomonas sp. DSM 50071 in the gut of superworms, highlighting a potential solution for PS biodegradation.

A novel endoscopic fluorescent band ligation method for tumor localization.

BACKGROUND: Accurate tumor localization is essential for minimally invasive surgery. This study describes the development of a novel endoscopic fluorescent band ligation method for the rapid and accurate identification of tumor sites during surgery. METHODS AND MATERIALS: The method utilized a fluorescent rubber band, made of indocyanine green (ICG) and a liquid rubber solution mixture, as well as a near-infrared fluorescence laparoscopic system with a dual light source using a high-powered light-emitting diode (LED) and a 785-nm laser diode. The fluorescent rubber bands were endoscopically placed on the mucosae of porcine stomachs and colons. During subsequent conventional laparoscopic stomach and colon surgery, the fluorescent bands were assayed using the near-infrared fluorescence laparoscopy system. RESULTS: The locations of the fluorescent clips were clearly identified on the fluorescence images in real time. The system was able to distinguish the two or three bands marked on the mucosal surfaces of the stomach and colon. Resection margins around the fluorescent bands were sufficient in the resected specimens obtained during stomach and colon surgery. CONCLUSION: These novel endoscopic fluorescent bands could be rapidly and accurately localized during stomach and colon surgery. Use of these bands may make possible the excision of exact target sites during minimally invasive gastrointestinal surgery.

Identification of plastic-degrading bacteria in the human gut.

Environmental pollution caused by the excessive use of plastics has resulted in the inflow of microplastics into the human body. However, the effects of microplastics on the human gut microbiota still need to be better understood. To determine whether plastic-degrading bacteria exist in the human gut, we collected the feces of six human individuals, did enrichment cultures and screened for bacterial species with a low-density polyethylene (LDPE) or polypropylene (PP)-degrading activity using a micro-spray method. We successfully isolated four bacterial species with an LDPE-degrading activity and three with a PP-degrading activity. Notably, all bacterial species identified with an LDPE or PP-degrading activity were opportunistic pathogens. We analyzed the microbial degradation of the LDPE or PP surface using scanning electron microscopy and confirmed that each bacterial species caused the physical changes. Chemical structural changes were further investigated using X-ray photoelectron spectroscopy and Fourier-transform-infrared spectroscopy, confirming the oxidation of the LDPE or PP surface with the formation of carbonyl groups (C=O), ester groups (CO), and hydroxyl groups (-OH) by each bacterial species. Finally, high temperature gel permeation chromatography (HT-GPC) analysis showed that these bacterial species performed to a limited extent depolymerization. These results indicate that, as a single species, these opportunistic pathogens in the human gut have a complete set of enzymes and other components required to initiate the oxidation of the carbon chains of LDPE or PP and to degrade them. Furthermore, these findings suggest that these bacterial species can potentially biodegrade and metabolize microplastics in the human gut.

Cesium Lead Bromide (CsPbBr3) Perovskite Quantum Dot-Based Photosensor for Chemiluminescence Immunoassays.

Chemiluminescence immunoassays have been widely employed for diagnosing various diseases. However, because of the extremely low intensity chemiluminescence signals, highly sensitive transducers, such as photomultiplier tubes and image sensors with cooling devices, are required to overcome this drawback. In this study, a hypersensitive photosensor was developed based on cesium lead bromide (CsPbBr3) perovskite quantum dots (QDs) with sufficient high sensitivity for chemiluminescence immunoassays. First, CsPbBr3 QDs with a highly uniform size, that is, 5 nm, were synthesized under thermodynamic control to achieve a high size confinement effect. For the fabrication of the photosensor, MoS2 nanoflakes were used as an electron transfer layer and heat-treated at an optimum temperature. Additionally, a parylene-C film was used as a passivation layer to improve the physical stability and sensitivity of the photosensor. In particular, the trap states on the CsPbBr3 QDs were reduced by the passivation layer, and the sensitivity was increased. Finally, a photosensor based on CsPbBr3 QDs was employed in chemiluminescence immunoassays for the detection of human hepatitis B surface antigen, human immunodeficiency virus antibody, and alpha-fetoprotein (AFP, a cancer biomarker). When compared with the conventionally used equipment, the photosensor was determined to be feasible for application in chemiluminescence immunoassays.

Rapid biodegradation of polyphenylene sulfide plastic beads by Pseudomonas sp.

Pseudomonas sp. isolated from soil, are bioremediating microorganisms that are capable of degrading various types of plastics. Polyphenylene sulfide (PPS) has the most excellent structural stability among general plastics and thus is extremely difficult to break down using physical or chemical methods. This study demonstrates the efficient biodegradation of PPS by Pseudomonas sp., which exists in the gut of superworms. Compared with the conventional film-type of plastic, the degradation efficiencies to the bead form of plastic were significantly improved and thus the biodegradation time was dramatically shortened. Therefore, instead of film-type plastics, we used 300 µm diameter plastic beads for the measurement of Pseudomonas sp.-mediated biodegradation of PPS during a 10-day period. This method not only can be used for comparison and verification of the biodegradation efficiency of different types of plastics within a short reaction time of 10 days, but also provides the possibility to develop a new and more efficient screening system to rapidly identify the most efficient species of bacteria for the biodegradation of various types of plastics.

Highly sensitive in situ-synthesized cadmium sulfide (CdS) nanowire photosensor for chemiluminescent immunoassays.

Highly sensitive in situ-synthesized cadmium sulfide (CdS) nanowires (NWs) for the detection of chemiluminescence in immunoassays with a photoresist (PR) layer to stabilize the CdS NWs before and after coating with a parylene film were developed. The thickness of the PR layer was controlled by adjusting the viscosity of the PR solution used for spin-coating. PR2005 was the optimal PR for passivation of the NW surface. After the addition of a parylene coating on the CdS NWs, the photocurrent increased by as much as 50% over a broad range of light intensities, and the additional PR layer increased the photoresponse over the whole range of light intensities. When the photoresponses of the CdS NWs with and without the parylene film were compared after the addition of a PR layer, significant differences were observed in the photocurrent behavior after the incident light was turned off. For the CdS NWs with a parylene film and PR layer, the photocurrent reached the baseline within milliseconds of the incident light being turned off. However, the CdS NWs without a parylene film but with a PR layer required >60 s to reach the baseline level. This difference was due to the capacitance arising from the contact between the NWs. The in situ-synthesized CdS NW photosensor passivated by the parylene film and a PR layer was used in a chemiluminescence-based immunoassay. Finally, the detection of human immunodeficiency virus antibodies was demonstrated via a chemiluminescent enzyme-linked immunosorbent assay based on the CdS NW photosensor in comparison with the optical-density measurement for the chromogenic reaction of TMB(3,3',5,5'-Tetramethylbenzidine).

Three-Dimensional Paper-Based Microfluidic Analytical Devices Integrated with a Plasma Separation Membrane for the Detection of Biomarkers in Whole Blood.

Paper-based microfluidic analytical devices (µPADs) have recently attracted attention as a point-of-care test kit because of their low cost and nonrequirement for external forces. To directly detect biomarkers in whole blood, however, they need to be assembled with a filter such as a plasma separation membrane (PSM) because the color of the blood cells interferes with the colorimetric assay. However, this assembly process is rather complicated and cumbersome, and the fluid does not uniformly move to the detection zone when the adhesion between the paper and PSM is not perfect. In this study, we report a simple three-dimensional (3D) printing method for fabricating PSM-integrated 3D-µPADs made of plastics without the need for additional assembly. In detail, PSM was coated with parylene C to prevent its dissolution from organic solvent during 3D printing. Then, the coated PSM was superimposed on the paper. Detection zones and a reservoir were printed on the paper and PSM via liquid photopolymerization, using a digital light processing printer. The limit of detection of the PSM-integrated 3D-µPADs for glucose in whole blood was 0.3 mM, and these devices demonstrated clinically relevant performance on diabetes patient blood samples. Our 3D-µPADs can also simultaneously detect multiple metabolic disease markers including glucose, cholesterol, and triglycerides in whole blood. Our results suggest that our printing method is useful for fabricating 3D-µPADs integrated with PSM for the direct detection of biomarkers in whole blood.

In situ-synthesized cadmium sulfide nanowire photosensor with a parylene passivation layer for chemiluminescent immunoassays.

The direct in situ synthesis of cadmium sulfide (CdS) nanowires (NWs) was presented by direct synthesis of CdS NWs on the gold surface of an interdigitated electrode (IDE). In this work, we investigated the effect of a strong oxidant on the surfaces of the CdS NWs using X-ray photoelectron spectroscopy, transmission electron microscopy, and time-of-flight secondary ion mass spectrometry. We also fabricated a parylene-C film as a surface passivation layer for in situ-synthesized CdS NW photosensors and investigated the influence of the parylene-C passivation layer on the photoresponse during the coating of parylene-C under vacuum using a quartz crystal microbalance and a photoanalyzer. Finally, we used the in situ-synthesized CdS NW photosensor with the parylene-C passivation layer to detect the chemiluminescence of horseradish peroxidase and luminol and applied it to medical detection of carcinoembryonic antigen.