Ni-rGO Sensor Combined with Human Olfactory Receptor-Embedded Nanodiscs for Detecting Gas-Phase DMMP as a Simulant of Nerve Agents.

Nerve agents are organophosphorus toxic chemicals that can inhibit acetylcholinesterase, leading to paralysis of the nervous system and death. Early detection of nerve agents is important for safety issues. Dimethyl methylphosphonate (DMMP) is widely used as a simulant of nerve agents, and many studies have been conducted using DMMP as a substitute for detecting nerve agents. Despite many studies on sensors for detecting DMMP, they have limitations in sensitivity and selectivity. To overcome these limitations, a nickel-decorated reduced graphene oxide (Ni-rGO) sensor with human olfactory receptor hOR2T7 nanodiscs was utilized to create a bioelectronic nose platform for DMMP gas detection. hOR2T7 was produced and reconstituted into nanodiscs for enhancing the sensor's stability, especially for detection in a gas phase. It could detect DMMP gas selectively and repeatedly at a concentration of 1 ppb. This sensitive and selective bioelectronic nose can be applied as a practical tool for the detection of gaseous chemical warfare agents in military and safety fields.

Multiscale pore contained carbon nanofiber-based field-effect transistor biosensors for nesfatin-1 detection.

Nesfatin-1 (NES1) is a potential biomarker found in serum and saliva that indicates hyperpolarization and depolarization in the hypothalamic ventricle nucleus as well as an increase in epileptic conditions. However, real-time investigations have not been carried out to detect changes in the concentration of NES1. In this study, we develop a multiscale pore contained carbon nanofiber-based field-effect transistor (FET) biosensor to detect NES1. The activated multiscale pore contained carbon nanofiber (a-MPCNF) is generated using a single-nozzle co-electrospinning method and a subsequent steam-activation process to obtain a signal transducer and template for immobilization of bioreceptors. The prepared biosensor exhibits a high sensitivity to NES1. It can detect levels as low as 0.1 fM of NES1, even in the presence of other interfering biomolecules. Furthermore, the a-MPCNF-based FET sensor has significant potential for practical applications in non-invasive real-time diagnosis, as indicated by its sensing performance in artificial saliva.

Facile Synthesis of Co3O4-Incorporated Multichannel Carbon Nanofibers for Electrochemical Applications.

Considering their superior electrochemical performances, extensive studies have been carried out on composite nanomaterials based on porous carbon nanofibers. However, the introduction of inorganic components into a porous structure is complex and has a low yield. In this study, we propose a simple synthesis of cobalt-oxide-incorporated multichannel carbon nanofibers (P-Co-MCNFs) as electrode materials for electrochemical applications. The cobalt oxide component is directly formed in the carbon structure by a simple oxygen plasma exposure of the phase-separated polymer nanofibers. P-Co-MCNF displays high specific capacitance (815 F g-1 at 2.0 A g-1), rate capability (821 F g-1 at 1 A g-1 and 786 F g-1 at 20 A g-1), and cycle stability (92.1% for 5000 cycles) as a supercapacitor electrode. Moreover, excellent sensitivity (down to 1 nM) and selectivity to the glucose molecule is demonstrated for nonenzyme sensor applications.

Mimicking cotranslational folding of prosubtilisin E in vitro.

Nascent polypeptides are synthesized on ribosomes starting at the N-terminus and simultaneously begin to fold during translation. We constructed N-terminal fragments of prosubtilisin E containing an intramolecular chaperone (IMC) at N-terminus to mimic cotranslational folding intermediates of prosubtilisin. The IMC-fragments of prosubtilisin exhibited progressive enhancement of their secondary structures and thermostabilities with increasing polypeptide length. However, even the largest IMC-fragment with 72 residues truncated from the C-terminus behaved as a molten globule, indicating the requirement of the C-terminal region to have a stable tertiary structure. Furthermore, truncation of the IMC in the IMC-fragments resulted in aggregation, suggesting that the IMC plays a crucial role to prevent misfolding and aggregation of cotranslational folding intermediates during translation of prosubtilisin polypeptide.

Fluorine plasma treatment on carbon-based perovskite solar cells for rapid moisture protection layer formation and performance enhancement.

A fluorine plasma-treated carbon electrode is used in HTM-free perovskite solar cells for high efficiency and moisture resistance. The fluorine-treated device with a champion power conversion efficiency (PCE) of 14.86% is achieved with a highly enhanced FF (FF = 0.69), showing superior long-term stability and excellent moisture penetration suppression.

Envelope-deforming antiviral peptide derived from influenza virus M2 protein.

Molecules interfering with lipid bilayer function exhibit strong antiviral activity against a broad range of enveloped viruses, with a lower risk of resistance development than that for viral protein-targeting drugs. Amphipathic peptides are rich sources of such membrane-interacting antivirals. Here, we report that influenza viruses were effectively inactivated by M2 AH, an amphipathic peptide derived from the M2 protein of the influenza virus. Although overall hydrophobicity () of M2 AH was not related to antiviral activity, modification of the hydrophobic moment (<µH>) of M2 AH dramatically altered the antiviral activity of this peptide. M2 MH, a derivative of M2 AH with a <µH> of 0.874, showed a half maximal inhibitory concentration (IC50) of 53.3 nM against the A/PR/8/34 strain (H1N1), which is 16-times lower than that of M2 AH. The selectivity index (IC50/CC50), where CC50 is the half maximal cytotoxic concentration, was 360 for M2 MH and 81 for M2 AH. Dynamic light scattering spectroscopy and electron microscopy revealed that M2 AH-derived peptides did not disrupt liposomes but altered the shape of viruses. This result suggests that the shape of virus envelope was closely related to its activity. Thus, we propose that deforming without rupturing the membranes may achieve a high selectivity index for peptide antivirals.

Delphinidin Chloride and Its Hydrolytic Metabolite Gallic Acid Promote Differentiation of Regulatory T cells and Have an Anti-inflammatory Effect on the Allograft Model.

Regulatory T cells (Tregs) control the reactivity of other T cells to prevent excessive inflammatory responses. They also plays a role in preventing autoimmune diseases; but when they are overproduced, they decreased vital immunity, which can lead to invasion of external pathogens. Therefore, it is most important in preventing the development of immune diseases to maintain the homeostasis of these cells. Delphinidin chloride is an anthocyanidin and known to have anti-oxidant activities. However, its structure is very unstable and easily decomposed. One of these degradation products is gallic acid, which also has anti-oxidant effects. In this study, we examined the effect of these materials on Tregs in controlling immune response. It was found that these materials further promote differentiation into Tregs, and TGF-ß and IL-2 related signals are involved in this process. Furthermore, it was verified that a variety of immunosuppressive proteins were secreted more, and the function of induced Tregs was also increased. Finally, in the allograft model, we could find a decrease in activated T cells when these materials were treated because they increased differentiation into Tregs. Therefore, these two materials are expected to become new candidates for the treatment of diseases caused by excessive activation of immune cells, such as autoimmune diseases. PRACTICAL APPLICATION: Delphinidin, a kind of anthocyanin rich in pigmented fruits, and its hydrolytic metabolite, gallic acid, are known to have antimicrobial and anti-oxidant properties. In this experiment, it was shown that delphinidin and gallic acid had an effect of increasing the differentiation of regulatory T cells, and the effect of suppressing the function of memory T cells was also observed. Due to these functions, delphinidin and gallic acid might have the potential to be used as immune suppressive agents in organ transplant and autoimmune disease patients or be a model for food development associated with the immune system.

Ultrasensitive and Selective Organic FET-type Nonenzymatic Dopamine Sensor Based on Platinum Nanoparticles-Decorated Reduced Graphene Oxide.

Dopamine (DA), a catecholamine hormone, is an important neurotransmitter that controls renal and cardiovascular organizations and regulates physiological activities. Abnormal concentrations of DA cause unfavorable neuronal illnesses such as Parkinson's disease, schizophrenia, and attention deficit hyperactivity disorder/attention deficit disorder. However, the DA concentration is exceedingly low in patients and difficult to detect with existing biosensors. In this study, we developed an organic field-effect-transistor-type (OFET) nonenzyme biosensor using platinum nanoparticle-decorated reduced graphene oxide (Pt_rGO) for ultrasensitive and selective DA detection. The Pt_rGOs were fabricated by reducing GO aqueous solution-containing Pt precursors (PtCl4) with a chemical reducing agent. The Pt_rGOs were immobilized on a graphene substrate by π-π interactions and a conducting-polymer source-drain electrode was patterned on the substrate to form the DA sensor. The resulting OFET sensor showed a high sensitivity to remarkably low DA concentrations (100 × 10-18 M) and selectivity among interfering molecules. Good stability was expected for the OFET sensor because it was fabricated without an enzymatic receptor, and π-π conjugation is a part of the immobilization process. Furthermore, the OFET sensors are flexible and offer the possibility of wide application as wearable and portable sensors.

Translation-dependent mRNA cleavage by YhaV in Escherichia coli.

Many bacteria have toxin-antitoxin (TA) systems, where toxin gene expression inhibits their own cell growth. mRNA is one of the well-known targets of the toxins in the type II toxin-antitoxin systems. Here, we examined the ribosome dependency of the endoribonuclease activity of YhaV, one of the toxins in type II TA systems, on mRNA in vitro and in vivo. A polysome profiling assay revealed that YhaV is bound to the 70S ribosomes and 50S ribosomal subunits. Moreover, we found that while YhaV cleaves ompF and lpp mRNAs in a translation-dependent manner, they did not cleave the 5' untranslated region in primer extension experiments. From these results, we conclude that YhaV is a ribosome-dependent toxin that cleaves mRNA in a translation-dependent manner.

Wireless, Room Temperature Volatile Organic Compound Sensor Based on Polypyrrole Nanoparticle Immobilized Ultrahigh Frequency Radio Frequency Identification Tag.

Due to rapid advances in technology which have contributed to the development of portable equipment, highly sensitive and selective sensor technology is in demand. In particular, many approaches to the modification of wireless sensor systems have been studied. Wireless systems have many advantages, including unobtrusive installation, high nodal densities, low cost, and potential commercial applications. In this study, we fabricated radio frequency identification (RFID)-based wireless sensor systems using carboxyl group functionalized polypyrrole (C-PPy) nanoparticles (NPs). The C-PPy NPs were synthesized via chemical oxidation copolymerization, and then their electrical and chemical properties were characterized by a variety of methods. The sensor system was composed of an RFID reader antenna and a sensor tag made from a commercially available ultrahigh frequency RFID tag coated with C-PPy NPs. The C-PPy NPs were covalently bonded to the tag to form a passive sensor. This type of sensor can be produced at a very low cost and exhibits ultrahigh sensitivity to ammonia, detecting concentrations as low as 0.1 ppm. These sensors operated wirelessly and maintained their sensing performance as they were deformed by bending and twisting. Due to their flexibility, these sensors may be used in wearable technologies for sensing gases.

Heterologous Expression of Der Homologs in an Escherichia coli der Mutant and Their Functional Complementation.

UNLABELLED: The unique Escherichia coli GTPase Der (double Era-like GTPase), which contains tandemly repeated GTP-binding domains, has been shown to play an essential role in 50S ribosomal subunit biogenesis. The depletion of Der results in the accumulation of precursors of 50S ribosomal subunits that are structurally unstable at low Mg(2+) concentrations. Der homologs are ubiquitously found in eubacteria. Conversely, very few are conserved in eukaryotes, and none is conserved in archaea. In the present study, to verify their conserved role in bacterial 50S ribosomal subunit biogenesis, we cloned Der homologs from two gammaproteobacteria, Klebsiella pneumoniae and Salmonella enterica serovar Typhimurium; two pathogenic bacteria, Staphylococcus aureus and Neisseria gonorrhoeae; and the extremophile Deinococcus radiodurans and then evaluated whether they could functionally complement the E. coli der-null phenotype. Only K. pneumoniae and S Typhimurium Der proteins enabled the E. coli der-null strain to grow under nonpermissive conditions. Sucrose density gradient experiments revealed that the expression of K. pneumoniae and S Typhimurium Der proteins rescued the structural instability of 50S ribosomal subunits, which was caused by E. coli Der depletion. To determine what allows their complementation, we constructed Der chimeras. We found that only Der chimeras harboring both the linker and long C-terminal regions could reverse the growth defects of the der-null strain. Our findings suggest that ubiquitously conserved essential GTPase Der is involved in 50S ribosomal subunit biosynthesis in various bacteria and that the linker and C-terminal regions may participate in species-specific recognition or interaction with the 50S ribosomal subunit. IMPORTANCE: In Escherichia coli, Der (double Era-like GTPase) is an essential GTPase that is important for the production of mature 50S ribosomal subunits. However, to date, its precise role in ribosome biogenesis has not been clarified. In this study, we used five Der homologs from gammaproteobacteria, pathogenic bacteria, and an extremophile to elucidate their conserved function in 50S ribosomal subunit biogenesis. Among them, Klebsiella pneumoniae and Salmonella enterica serovar Typhimurium Der homologs implicated the participation of Der in ribosome assembly in E. coli Our results show that the linker and C-terminal regions of Der homologs are correlated with its functional complementation in E. coli der mutants, suggesting that they are involved in species-specific recognition or interaction with 50S ribosomal subunits.

Affinity improvement by fine tuning of single-chain variable fragment against aflatoxin B1.

Aflatoxin B1 (AFB1) produced in Aspergillus flavus is a major hepatocarcinogen found in foods and feed. For effective immunological detection of AFB1 at low concentrations, the development of high affinity antibody for AFB1 is required. Previously, an affinity-maturated single-chain variable fragment containing 6 mutations (scFv-M37) was isolated from an artificial mutagenic library, which showed a 9-fold higher affinity than its wild type scFv. In this study, the effect of the 6 mutated residues on the affinity improvement was characterized using surface plasmon resonance analysis, which identified a deleterious mutation (VH-A110T) located on a framework region of the scFv-M37. The back mutation of VH-A110T resulted in a 3.2-fold affinity improvement, which was attributed to decrease of dissociation rate constant (kd) in interaction between AFB1 and the back mutant scFv. The biophysical analyses using circular dichroism and gel filtration revealed that the back mutation of VH-A110T caused a subtle conformational change of the scFv toward tighter binding to AFB1.

Ultrasensitive Bisphenol A Field-Effect Transistor Sensor Using an Aptamer-Modified Multichannel Carbon Nanofiber Transducer.

Bisphenol A (BPA) is a known endocrine-disrupting compound (EDC) that has a structure similar to that of the hormone estrogen. Even low concentrations of BPA are able to bind estrogen receptors, thereby inducing severe diseases such as reproductive disorders, chronic diseases, and various types of cancer. Despite such serious effects, the use of BPA remains widespread. Therefore, monitoring of both dietary and nondietary exposure to BPA is important for human healthcare. Herein, we present a field-effect transistor (FET) sensor using aptamer-modified multichannel carbon nanofibers (MCNFs) to detect BPA. The MCNFs are fabricated via single-nozzle electrospinning of two immiscible polymer solutions followed by thermal treatment in an inert atmosphere. The MCNFs are then oxidized using a solution of HNO3 and H2SO4 to introduce carboxyl groups on the surface of the fibers. The carboxyl-functionalized MCNFs (CMCNFs) are immobilized on an amine-functionalized electrode substrate by forming a covalent bond, and amine-functionalized BPA-binding aptamers are modified in the same manner on the CMCNFs. The resulting FET sensors exhibit a high sensitivity, as well as specificity toward BPA at an unprecedentedly low concentration of 1 fM. Furthermore, these sensors are stable and could be reused for repeated assays.

Porous palladium coated conducting polymer nanoparticles for ultrasensitive hydrogen sensors.

Hydrogen, a clean-burning fuel, is of key importance to various industrial applications, including fuel cells and in the aerospace and automotive industries. However, hydrogen gas is odorless, colorless, and highly flammable; thus appropriate safety protocol implementation and monitoring are essential. Highly sensitive hydrogen leak detection and surveillance sensor systems are needed; additionally, the ability to maintain uniformity through repetitive hydrogen sensing is becoming increasingly important. In this report, we detail the fabrication of porous palladium coated conducting polymer (3-carboxylate polypyrrole) nanoparticles (Pd@CPPys) to detect hydrogen gas. The Pd@CPPys are produced by means of facile alkyl functionalization and chemical reduction of a pristine 3-carboxylate polypyrrole nanoparticle-contained palladium precursor (PdCl(2)) solution. The resulting Pd@CPPy-based sensor electrode exhibits ultrahigh sensitivity (0.1 ppm) and stability toward hydrogen gas at room temperature due to the palladium sensing layer.

Multidimensional MnO2 nanohair-decorated hybrid multichannel carbon nanofiber as an electrode material for high-performance supercapacitors.

One-dimensional (1D)-structured nanomaterials represent one of the most attractive candidates for energy-storage systems due to their contribution to design simplicity, fast charge-transportation network, and their allowance for more accessible ion diffusion. In particular, 1D-structured nanomaterials with a highly complex inner-pore configuration enhance functionality by taking advantage of both the hollow and 1D structures. In this study, we report a MnO2 nanohair-decorated, hybrid multichannel carbon nanofiber (Mn_MCNF) fabricated via single-nozzle co-electrospinning of two immiscible polymer solutions, followed by carbonization and redox reactions. With improved ion accessibility, the optimized Mn_MCNF sample (Mn_MCNF_60 corresponding to a reaction duration time of 60 min for optimal MnO2 nanohair growth) exhibited a high specific capacitance of 855 F g(-1) and excellent cycling performance with ∼87.3% capacitance retention over 5000 cycles.

Flower-like Palladium Nanoclusters Decorated Graphene Electrodes for Ultrasensitive and Flexible Hydrogen Gas Sensing.

Flower-like palladium nanoclusters (FPNCs) are electrodeposited onto graphene electrode that are prepared by chemical vapor deposition (CVD). The CVD graphene layer is transferred onto a poly(ethylene naphthalate) (PEN) film to provide a mechanical stability and flexibility. The surface of the CVD graphene is functionalized with diaminonaphthalene (DAN) to form flower shapes. Palladium nanoparticles act as templates to mediate the formation of FPNCs, which increase in size with reaction time. The population of FPNCs can be controlled by adjusting the DAN concentration as functionalization solution. These FPNCs_CG electrodes are sensitive to hydrogen gas at room temperature. The sensitivity and response time as a function of the FPNCs population are investigated, resulted in improved performance with increasing population. Furthermore, the minimum detectable level (MDL) of hydrogen is 0.1 ppm, which is at least 2 orders of magnitude lower than that of chemical sensors based on other Pd-based hybrid materials.

Inhibitory effect of UvrD and DinG on the replication of ColE1-derived plasmids in Escherichia coli.

CspA has been identified as a major cold-shock protein in Escherichia coli. CspA binds to RNAs which are abnormally folded at low temperature and then acts as an RNA chaperone unfolding those RNAs. The dramatic expression of cspA at low temperature is contributed by posttranscriptional stability and robust translatability. Interestingly, when cspA mRNA encoding a premature nonsense codon was overexpressed at low temperature, cell growth was completely inhibited. This phenotype was termed LACE (the low temperature-dependent antibiotic effect of truncated cspA expression), and this lethality resulted from exclusive stalling of most ribosomes on mutant cspA mRNAs. In a previous study, we demonstrated that overexpression of the ATP-dependent DNA helicases, UvrD and DinG, suppressed the lethality and ribosome stalling caused by mutant cspA mRNA. In the present study, we attempted to elucidate how these two DNA helicases help recover normal growth under LACE condition. Interestingly, we found that UvrD and DinG appeared to have an ability to down-regulate the replication of pUC-based high copy plasmid. In plasmid copy number tests, the amount of pUC-based plasmid encoding mutant cspA was reduced by 3-10-fold when either UvrD or DinG was expressed. Through a ß-galactosidase activity assay, we also confirmed that expression of the lacZα gene inserted into the pUC-based plasmid was significantly reduced due to down-regulation of plasmid replication. Our findings imply that UvrD and DinG, known as non-replicative helicases, play a novel role in the regulation of ColE1-like plasmid replication.

Affinity maturation of single-chain variable fragment specific for aflatoxin B(1) using yeast surface display.

As aflatoxin B1 is one of the most toxic mycotoxins, it is important to detect and to quantify aflatoxin B1 accurately by immunological methods. To enhance aflatoxin B1-binding affinity of the single-chain variable fragment, yeast surface display technique combined with fluorescence-activated cell sorting was applied. A randomly mutated scFv library was subjected to 4 rounds of fluorescence-activated cell sorting, resulting in isolation of 5 scFv variants showing an affinity improvement compared to the parental wild type scFv. The best scFv with a 9-fold improvement in affinity for aflatoxin B1 exhibited similar specificity to the monoclonal antibody. Most of the mutations in scFv-M37 were located outside of the canonical antigen-contact loops, suggesting that its affinity improvement might be driven by an allosteric effect inducing scFv-M37 to form a more favorable binding pocket for aflatoxin B1 than the wild type scFv.

Highly Sensitive and Selective Field-Effect-Transistor NonEnzyme Dopamine Sensors Based on Pt/Conducting Polymer Hybrid Nanoparticles.

Dopamine (DA), as one of catecholamine family of neurotransmitters, is crucially important in humans owing to various critical effects on biometric system such as brine circuitry, neuronal plasticity, organization of stress responses, and control of cardiovascular and renal organizations. Abnormal level of dopamine in the central nervous system causes several neurological diseases, e.g., schizophrenia, Parkinson's disease, and attention deficit hybperactivity disorder (ADHD)/attention deficit disorder (ADD). In this report, we suggest the fabrication of nonenzyme field effect transistor (FET) sensor composed of immobilized Pt particle decorated conducting-polymer (3-carboxylate polypyrrole) nanoparticles (Pt_CPPy) to detect dopamine. The hybrid nanoparticles (NPs) are produced by means of facile chemical reduction of pristine CPPyNP-contained Pt precursor (PtCl4 ) solution. The Pt_CPPys are then immobilized on an amine-functionalized (-NH2 ) interdigitated-array electrode substrate, through the formation of covalent bonds with amine groups (-CONH). The resulting Pt_CPPy-based FET sensors exhibit high sensitivity and selectivity toward DA at unprecedentedly low concentrations (100 × 10(-15) m) and among interfering biomolecules, respectively. Additionally, due to the covalent bonding involved in the immobilization process, a longer lifetime is expected for the FET sensor.

Detection of hazardous gas using multidemensional porous iron oxide nanorods-decorated carbon nanoparticles.

Multidimensional porous iron oxide (Fe2O3) nanorods-decorated carbon nanoparticles (MPFCNPs) were fabricated using a dual-nozzle electrospray, thermal stirring, and heat treatment. Polypyrrole (PPy) NPs with FeOOH nanorods were synthesized by electrospraying Fe(3+) ions, which were adsorbed on the PPy NP surface; the adsorbed Fe(3+) ions reacted with NaOH to create FeOOH nuclei, and then followed thermal stirring grow nanorods without aggregation. MPFCNPs were fabricated through heat treatment, with the porous structure created in the Fe2O3 nanorods by hydroxyl group decomposition. The size of the MPFCNPs and the length of the porous Fe2O3 nanorods were controlled by the PPy NP template and concentration of initiator solution, respectively. The MPFCNPs were then utilized as a chemical sensor transducer for NO2 gas detection at room temperature. The response of the MFPCNP sensor was highly sensitive, displaying a minimum detectable level of 1 ppm; this detection level is lower than that of organic-inorganic hybrid sensors. Moreover, sensitivity also improved with decreasing the diameter of MPFCNPs and increasing Fe2O3 nanorod length. The enhanced sensitivity was attributed to the larger surface area presented by the particle size and the porous structure.