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A conceptually new on-site OTA detection in Baijiu is developed based on DNA base pair stacking assembly of gold nanoparticles.

Demonstration of a colorimetric approach for nucleic acid detection represents an attractive educational experiment for chemistry undergraduate students in the time of coronavirus pandemic. Herein, a rapid and vivid detection method that visualizes the presence of a specific DNA sequence is described. The plasmon resonance of gold nanoparticles (AuNPs) (∼20 nm in diameter) endows a red color to DNA-functionalized gold nanoparticles (DNA-AuNPs) in a colloidal solution. Simply upon addition of a sample containing the complementary DNA sequence, the DNA-AuNPs dispersed in a strong ionic solution can spontaneously aggregate in minutes and turn to be purple in color, which can be detected using UV–visible spectroscopy as well as the naked eye. In contrast, the DNA-AuNPs remain highly dispersed as a reddish colloid even at high ionic strength when the DNA sequence is terminally mismatched or overhung. According to the teaching experience, this DNA testing experiment can be accomplished by students in pairs or groups under instruction in a three-day lab session. Given the rapid and fail-proof DNA testing, the testing method is also well-suited for hands-on introduction of analytical chemistry and nanotechnology to chemistry freshman and high school students.

A colorimetric assay of DNA cleavage by bleomycin (BLM) derivatives was developed utilizing high colloidal stability on double-stranded (ds) DNA-modified gold nanoparticles (dsDNA-AuNPs) possessing a cleavage site. The assay was performed using dsDNA-AuNPs treated with inactive BLM or activated BLM (Fe(II)⋅BLM). A 10-min exposure in dsDNA-AuNPs with inactive BLM treatment resulted in a rapid color change from red to purple because of salt-induced non-crosslinking aggregation of dsDNA-AuNPs. In contrast, the addition of active Fe(II)⋅BLM retained the red color, probably because of the formation of protruding structures at the outermost phase of dsDNA-AuNPs caused by BLM-mediated DNA cleavage. Furthermore, the results of our model experiments indicate that oxidative base release and DNA-cleavage pathways could be visually distinguished with color change. The present methodology was also applicable to model screening assays using several drugs with different mechanisms related to antitumor activity. These results strongly suggest that this assay with a rapid color change could lead to simple and efficient screening of potent antitumor agents.

Quality control of food products requires the development of powerful analytical tools and regulations to avoid fraud to consumers and ineffective product recalls. DNA barcoding represents a potent...

Self-assembly performed in ionic liquids (ILs) as a unique solvent promises distinct functions and applications in sensors, therapeutics, and optoelectronic devices due to the rich interactions between nanoparticle building blocks and ILs. However, the general consideration that common nanoparticles are readily destabilized by counterions in an IL has largely prevented researchers from investigating controlled nanoparticle assembly in IL-based systems. This study explores the assembling behaviour of double-stranded (ds) DNA-functionalized gold nanoparticles (dsDNA-AuNPs) in hydrated ionic liquids. The DNA base pair stacking assembly of dsDNA-AuNPs occurs at a low IL concentration (5%). However, a moderate ionic liquid concentration (5-40%) can de-hybridize dsDNA and leaves single-stranded (ss) DNA stabilizing the AuNPs. In concentrated ionic liquids (40%), interestingly, the higher ionic strength leads to the assembly of DNA-AuNPs. The triphasic assembly trend is also generally observed regardless of the type of IL. By down-regulation of DNA's melting temperature with the IL, the assembly of DNA-AuNPs affords robust response to a lower temperature range, promising applications in plasmonic devices and range-tunable temperature sensors.

Smart thermal switching of plasmonic materials holds important implications for photothermal therapy, optoelectronics, and information storage. Due to the difficulty in translation of interplays of interfacial molecular behaviors to highly controlled interparticle spacing, unfortunately, realization of a plasmon switch with rapid, sensitive, and tunable thermo-response remains challenging. We report the development of a smart plasmon switch based on gold nanorods (AuNRs) through terminal breathing of surface-grafted double-stranded (ds) DNA in alcoholic solvents. By closing dsDNA terminals on the AuNRs, DNA base pair stacking assembly of the AuNRs occurs immediately, whereas dispersed AuNRs can be restored at an elevated temperature owing to the opening of the dsDNA ends. Both electron microscopic characterizations and optical simulations confirmed that the DNA terminal breathing behavior has a significant impact on the interfacial interactions of the AuNRs and brings about repeated plasmon switching that exhibits a thermal sensitivity of 1 °C. Importantly, by altering the composition of water–alcohol mixtures, the plasmon switch can be continuously tuned with respect to temperature response range while preserving excellent switching performance. Given the high sensitivity and wide response range, the bioinspired plasmonic materials suggest expanded application scope.

This study was performed to determine the risk factors associated with postoperative complications after surgery under general anesthesia according to respiratory function test results and oral conditions. Preoperative examination data were collected for 471 patients who underwent surgery under general anesthesia at the Medical Hospital of Kyusyu University. Respiratory function tests, oral examinations, and perioperative oral management were performed in all patients. The incidence of and risk factors for postoperative complications were investigated. Classification and regression tree analyses were performed to investigate the risk factors for postoperative complications. Among the 471 patients, 11 developed postoperative pneumonia, 10 developed postoperative respiratory symptoms, and 10 developed postoperative fever. The most important risk factor for pneumonia was edentulism. Age, the Brinkman index, and head and neck surgery were also revealed as important risk factors for pneumonia. The O’Leary plaque control record (initial visit) was an important risk factor for postoperative respiratory symptoms. With respect to postoperative fever, a Hugh–Jones classification of grade > 1 was the most important risk factor; edentulism and a Brinkman index of > 642.5 were also found to be risk factors. In addition to respiratory function tests, oral examinations may be important for the prediction of postoperative complications. Additionally, improved oral hygiene may be effective in preventing postoperative respiratory complications. Risk factors for postoperative complications should be comprehensively evaluated using both respiratory function tests and oral findings.

Hierarchical plasmonic nanostructures that embed multiple shape-discrete domains and thereby unique plasmon resonance have implications in detection, diagnostics, and photovoltaics. Despite great progress in self-assembly of gold nanostructures, direct synthesis of hierarchical Au nanostructures that afford dense and steady intrinsic hotspots has remained less explored. This is partially because nanocrystal growth principally yields symmetrical nanocrystals with simple morphologies in monometallic systems. We report on the seeded synthesis of “Au nanograsses” on Au nanoplates that comprise an array of bent Au nanowire and exhibit total light absorption and intrinsically coupled plasmon resonance. The growth of Au nanograsses is achieved through competition between Au deposition and surface ligand passivation, which restricts the deposition of upcoming Au atoms onto the freshly deposited Au surfaces that have not been passivated. By controlling reaction kinetics and ligand passivation, the secondary nanostructures can be tuned from nanowires to nanotubes and near-spherical islands. Theoretical simulations indicate a stronger near-field electromagnetic field and a denser distribution of built-in hotspots of the Au nanograss than the near-spherical nanoisland array. Application of the Au nanograss in single-particle surface-enhanced Raman scattering (SERS) reveals a greatly elevated enhancement effect of the Au nanograss compared to the nanoisland array by nearly one order-of-magnitude.

Strains in biomolecules greatly restrict their structural flexibility. The effects of DNA's structural flexibility on nanoparticle stability have remained less explored in the field of plasmonic biosensors. In the present study, we discover the opposite effects of a rigid loop and a flexible single-stranded DNA (ssDNA) region in DNAzyme on the colloidal stability of gold nanoparticles (AuNPs), which afford "turn-on" plasmonic detection of Pb

The interactions between metal ions and biomolecules are crucial to various bioprocesses. Development of plasmon switching nanodevices that exploit these molecular interactions is of fundamental and technological interest. Here, we show plasmon switching based on rapid aggregation/dispersion of double-stranded DNA-modified gold nanorods (dsDNA–AuNRs) that exhibit colloidal behaviors depending on pairing/unpairing of the terminal bases. The dsDNA–AuNRs bearing a thymine–thymine (T–T) mismatch at the penultimate position undergo spontaneous non-cross-linking aggregation in the presence of Hg2+ due to T–Hg–T base pairing. Inversely, the subsequent addition of cysteine (Cys) gives rise to the removal of Hg2+ from the T–Hg–T base pair to reproduce the T–T mismatch, resulting in stable dispersion of the dsDNA–AuNRs. The chemical-responsive plasmon switch allows for the rapid and repeatable cycles at room temperature. The validity of the present method is further exemplified by developing another plasmon switch ...

A strategy is described to produce an anisotropic assembly of isotropic particles. To generate the anisotropy, local-structure-sensitive colloidal stability of double-stranded DNA-modified gold nanoparticles was exploited; namely, fully matched (F) particles are spontaneously aggregated at high ionic strength, whereas terminal-mismatched (M) particles continue to stably disperse. Linear trimers prepared by aligning both the F and M particles on a DNA template in a strictly defined order undergo highly directed assembly, as revealed by electron microscopy. Importantly, the identity of the central particle controls the structural anisotropy. The trimers containing the M or F particle at the center selectively assemble in an end-to-end or side-by-side manner, respectively. Further, similar trimers having a central M larger than the peripheral F form assemblies that have small particles between the large particles. By contrast, the trimers with a central F larger than the peripheral M form an assembled struct...

Au nanoplates represent an emerging class of plasmonic nanostructures of anisotropy that are promising great performance as a probe in biosensing and a building block for nanodevices. In spite of the rapid development of solution-phase synthesis, it has remained an open challenge for colloidal chemists to prepare Au nanoplates in high yield established for their Ag counterparts. This limitation largely compromises the practical use of Au nanoplates in plasmonic applications. Here we report a simple yet versatile approach for efficient purification of Au nanoplates in diverse colloidal mixtures based upon micelle-induced depletion flocculation. By means of calculation on free energy change upon depletion flocculation, we are able to pre-determine the flocculation conditions of different nanoparticles, allowing for predictable nanoparticle precipitation that depends on size and shape. We further demonstrate that the present approach is capable of isolating Au nanoplates of varied sizes in low yield (

Increasing prevalence of pollen allergies has raised concerns about human health. Development of a facile and precise method to detect pollen allergens would thus be of significance for environmental assessments and medical diagnoses. Here we report a sensitive colorimetric method to detect the Japanese cedar pollen allergen, Cry j 2. The method consists of two steps: a signal amplification based on the catalytic DNA hairpin self-assembly, followed by a signal transduction using the salt-induced non-cross-linking aggregation of gold nanoparticles densely modified with short DNA. The assay exhibits a detection limit of 0.2 ng/mL, which is 130-fold greater than that of the previously reported one. Moreover, the assay enables the detection of Cry j 2 spiked in soil solutions by avoiding any interference from the contaminants. The signal amplification system includes an anti-Cry j 2 DNA aptamer, which accounts for the absence of false responses to five nontarget allergen proteins. The present method could be of general applicability to various proteins by using appropriate aptamers.

Anisotropic gold nanoparticles have attracted great interest due to their unique physicochemical properties derived from the shape anisotropy. Manipulation of their interfacial interactions, and thereby the assembling behaviors are often requisite in their applications ranging from optical sensing and diagnosis to self-assembly. Recently, the control of interfacial force based on base pair stacking of DNA terminals have offered a new avenue to surface engineering of nanostructures. In this review, we focus on the DNA base stacking-induced assembly of anisotropic gold nanoparticles, such as nanorods and nanotriangles. The fundamental aspects of anisotropic gold nanoparticles are provided, including the mechanism of the anisotropic growth, the properties arising from the anisotropic shape, and the construction of DNA-grafted anisotropic gold nanoparticles. Then, the advanced applications of their functional assemblies in biosensing and ordered assembly are summarized, followed by a comparison with gold nanospheres. Finally, conclusions and the direction of outlooks are given including future challenges and opportunities in this field.

Gold nanoparticle (AuNP) assemblies in two-dimensions (2D) exhibit collective physical/chemical properties that are useful for various devices. However, technical issues still impede the efficient ordering of differently sized AuNPs on solid supports while avoiding phase separation. This paper describes a method to construct binary 2D assemblies by folding precursory circular chains composed of small and large AuNPs. The structural change is caused by a spontaneous, non-cross-linking assembly of fully matched double-stranded DNA-modified AuNPs (dsDNA-AuNPs) at a high ionic strength. Since larger dsDNA-AuNPs have a lower critical coagulation concentration of the supporting electrolyte, the spontaneous assembly of large AuNPs precedes that of small AuNPs in the precursory chain during evaporation. Transmission electron microscopy reveals that alternate-type AuNP chains are folded into a binary 2D structure in a mixed mode, whereas block-type chains are transformed into a binary 2D structure in a core-shell mode. The methodology could potentially be harnessed for the fabrication of binary AuNP arrays for various devices.

A colorimetric method for fast, simple, and selective detection of Cu2+ was developed using I–-mediated etching of triangular gold nanoplates (AuNPLs). The method was based on our finding that Cu2+ efficiently promoted this etching in the presence of SCN–. The etching process was accompanied by a dramatic color change from blue to red, allowing for visual and spectroscopic detection of Cu2+ with detection limits of 10 and 1 μM, respectively. When molecular recognition by a DNA aptamer was incorporated into this method, visual detection of chloramphenicol was also achieved with a detection limit of 5 μM.

As a class of emerging nanoparticles, gold nanotriangles (AuNTs) are characterized by unique structural anisotropy and plasmonic properties. The organization of AuNTs into well-defined architecture potentially promises collective properties that are difficult to produce by individual AuNTs. To date, however, the orientation-controlled self-assembly of AuNTs has been achieved with limited success. Here, we describe an effective and straightforward approach to induce directed self-assembly of AuNTs. By taking advantage of the uneven chemical reactivity of AuNT surfaces, we implement regioselective modification of the edges and the top/bottom surfaces with two different double-stranded DNA (dsDNA) sequences. By means of terminal single base pairing/unpairing, controlled assembly of the dsDNA-modified AuNTs evolves in a face-to-face or edge-to-edge manner based on blunt-end stacking interaction on an intentional region of the AuNTs, along with entropic repulsion by unpaired terminal nucleobases on the other region. This approach could be useful for achieving directed self-assembly of other anisotropic nanoparticles.

Involvement of single‐nucleotide polymorphism (SNP) genotyping in healthcare should allow for more effective use of pharmacogenomics. However, user‐friendly assays without the requirement of a special instrument still remain unavailable. This study describes naked‐eye SNP discrimination in exon 5 of the human cytochrome P450 2C19 monooxygenase gene, CYP2C19*1 (the wild‐type allele) and CYP2C19*2 (the variant allele with G681A point mutation). The present assay is composed of allele‐specific single‐base primer extension and salt‐induced aggregation of DNA‐modified gold nanoparticles (DNA–AuNPs). Genetic samples extracted from human hair roots are subjected to this assay. The results are verified by direct sequencing. This study should promise the prospective use of DNA–AuNPs in gene diagnosis.

Colloidal behaviors of particles functionalized with biomolecules are generally complicated. This study describes that colloidal behaviors of double-stranded (ds) DNA-carrying polymer micelles are well controlled by altering the molar ratio of single-stranded (ss) DNA moiety in the dsDNA shell. ssDNA-carrying micelles composed of a poly( N-isopropylacrylamide) (PNIPAAm) core surrounded by a dense shell of ssDNAs were prepared through self-assembly of PNIPAAm grafted with ssDNA by incubating its solution above the lower critical solution temperature. Spontaneous, non-cross-linking aggregation of the micelles was triggered by DNA duplex formation on the surface. Comparison of the critical coagulation concentration of NaCl among a series of the DNA-carrying micelles revealed the relationship between the helical structure of the surface-bound DNA and the colloidal stability of the micelles. The electrophoretic mobility analysis of the micelles indicated that the duplex formation reduced the structural flexibility of the surface-bound DNA, thereby decreasing the interparticle entropic repulsion. It is also suggested that the augmented rigidity of the surface-bound DNA increases the number of terminal base pairs facing the solvent, which could lead to multiple blunt-end stacking interaction among the micelles. Therefore, small DNA molecules could be considered unique surface-modifiers capable of controlling interactions between the surfaces of materials.

We have previously reported that a dynamic air-pressure sensor system allows respiratory status to be visually monitored for patients in minimally clothed condition. The dynamic air-pressure sensor measures vital information using changes in air pressure. To utilize this device in the field, we must clarify the influence of clothing conditions on measurement. The present study evaluated use of the dynamic air-pressure sensor system as a respiratory monitor that can reliably detect change in breathing patterns irrespective of clothing. Twelve healthy volunteers reclined on a dental chair positioned horizontally with the sensor pad for measuring air-pressure signals corresponding to respiration placed on the seat back of the dental chair in the central lumbar region. Respiratory measurements were taken under 2 conditions: (a) thinly clothed (subject lying directly on the sensor pad); and (b) thickly clothed (subject lying on the sensor pad covered with a pressure-reducing sheet). Air-pressure signals were recorded and time integration values for air pressure during each expiration were calculated. This information was compared with expiratory tidal volume measured simultaneously by a respirometer connected to the subject via face mask. The dynamic air-pressure sensor was able to receive the signal corresponding to respiration regardless of clothing conditions. A strong correlation was identified between expiratory tidal volume and time integration values for air pressure during each expiration for all subjects under both clothing conditions (0.840–0.988 for the thinly clothed condition and 0.867–0.992 for the thickly clothed condition). These results show that the dynamic air-pressure sensor is useful for monitoring respiratory physiology irrespective of clothing.

Self-assembled structures of metallic nanoparticles with dynamically changeable interparticle distance hold promise for the regulation of collective physical properties. This paper describes gold nanoparticle dimers and trimers that exhibit spontaneous and reversible changes in interparticle distance. To exploit this property, a gold nanoparticle is modified with precisely one long DNA strand and approximately five short DNA strands. The long DNA serves to align the nanoparticles on a template DNA via hybridization, while the short DNAs function to induce the interparticle distance changes. The obtained dimer and trimer are characterized with gel electrophoresis, dynamic light scattering measurements, and transmission electron microscopy (TEM). When the complementary short DNA is added to form the fully matched duplexes on the particle surface in the presence of MgCl2 , spontaneous reduction of the interparticle distance is observed with TEM and cryo-electron microscopy. By contrast, when the terminal-mismatched DNA is added, no structural change occurs under the same conditions. Therefore, the single base pairing/unpairing at the outermost surface of the nanoparticle impacts the interparticle distance. This unique feature could be applied to the regulation of structures and properties of various DNA-functionalized nanoparticle assemblies.

Directed assemblies of anisotropic metal nanoparticles exhibit attractive physical and chemical properties. However, an effective methodology to prepare differently directed assemblies from the same anisotropic nanoparticles is not yet available. Gold nanorods (AuNRs) region-selectively modified with different DNA strands can form side-by-side (SBS) and end-to-end (ETE) assemblies in a non-crosslinking manner. When the complementary DNA is hybridized to the surface-bound DNA, stacking interaction between the blunt ends takes place in the designated regions. Such AuNRs assemble into highly ordered structures, assisted by capillary forces emerging on the substrate surface. Moreover, insertion of a mercury(II)-mediated thymine-thymine base pair into the periphery of the DNA layer allows selective formation of the SBS or ETE assemblies from the strictly identical AuNRs with or without mercury(II).

A single-nucleotide polymorphism (SNP) detection method was developed by combining single-base primer extension and salt-induced aggregation of gold nanoparticles densely functionalized with double-stranded DNA (dsDNA-AuNP). The dsDNA-AuNPs undergo rapid aggregation in a medium of high ionic strength, whereas particles having a single-base protrusion at the outermost surface disperse stably, allowing detection of a single-base difference in length by color changes. When SNP typing primers are used as analytes to hybridize to the single-stranded DNA on the AuNP surface, the resulting dsDNA-AuNP works as a visual indicator of single-base extension. A set of four extension reaction mixtures is prepared using each of ddNTPs and subsequently subjected to the aggregation assay. Three mixtures involving ddNTP that is not complementary to the SNP site in the target produce the aggregates that exhibit a purple color. In contrast, one mixture with the complementary ddNTP generates the single-base protrusion and appears red. This method could potentially be used in clinical diagnostics for personalized medicine.

Temperature-responsive diblock copolymers made of poly(N-isopropylacrylamide) (PNIPAAm) generated by reversible addition-fragmentation chain transfer (RAFT) polymerization and a single-stranded DNA (ssDNA) self-assembled into polymer micelles. The micelles consisted of the PNIPAAm core surrounded by the ssDNA corona with a hydrodynamic diameter up to 300 nm in an aqueous medium above the lower critical solution temperature. In a medium of high ionic strength, the formation of the fully matched duplex with the complementary ssDNA on the surface of the polymer micelles induced rapid and spontaneous aggregation. By contrast, the micelles remained dispersed under the identical conditions when single-base-substituted ssDNA was added to form the corresponding terminal-mismatched duplex on the micellar surface. This highly sequence-selective process took place irrespective of the size of the PNIPAAm core.

To quantitatively detect single-base mutations in genetic samples, free-solution affinity capillary electrophoresis was developed using a diblock copolymer probe composed of allele-specific oligode...

Gold nanoparticles densely modified with single-stranded DNA (ssDNA-AuNPs) form aggregates with cross-linker ssDNAs via duplex formation. Alternatively, the ssDNA-AuNPs are spontaneously aggregated at high ionic strength in a non-cross-linking manner when complementary ssDNAs are added to form fully matched duplexes. Both aggregation modes are accompanied by a red-to-purple color change, which has been exploited in various bioassays. The current study compares the rapidity of color change between the cross-linking and non-cross-linking aggregation modes under identical conditions. When a small number of cross-linker/complementary DNAs are provided, the cross-linking mode exhibited more rapid color change than the non-cross-linking mode. Conversely, with a large number of the DNAs, the non-cross-linking aggregation occurred more rapidly than the cross-linking counterpart. This finding allows one to select a more appropriate aggregation mode for application of ssDNA-AuNPs to colorimetric assays under given conditions.

An affinity capillary electrophoresis method was developed to determine a binding constant between a peptide nucleic acid (PNA) and a hairpin-structured DNA. A diblock copolymer composed of PNA and polyethylene glycol (PEG) was synthesized as a novel affinity probe. The base sequence of the probe's PNA segment was complementary to a hairpin-structured region of a 60-base single-stranded DNA (ssDNA). Upon applying a voltage, the DNA hairpin migrated slowly compared to a random sequence ssDNA in the presence of the PNA probe. This retardation was induced by strand invasion of the PNA into the DNA hairpin to form a hybridized complex, where the PEG segment received a large amount of hydrodynamic friction during electrophoresis. The binding constant between the PNA probe and the DNA hairpin was easily determined by mobility analysis. This simple method would be potentially beneficial in studying binding behaviors of various artificial nucleotides to natural DNA or RNA.

Poly(N-isopropylacrylamide) (PNIPAAm) grafted with single-stranded (ss) DNA conjugate (PNIPAAm-g-DNA) self-assembles above its lower critical solution temperature to form colloidal particles. When the ssDNA within the particle hybridizes with its complementary DNA, the particles aggregate above a certain threshold of salt concentration with drastically increased turbidity in solution. Detailed structural information of the particle was obtained mainly by small-angle X-ray scattering. The influence of copolymer composition on the morphology of particle and non-crosslinking aggregation was examined. The particle consists of hydrophobic PNIPAAm core surrounded by hydrophilic DNA strands. The increase in DNA fraction brought about a significant decrease in core size, whereas the shell thickness little changed and corresponded to the length of DNA. A structural model with a sticky potential was applied to the analysis of particle aggregate. This analysis provided that the particles aggregate while the coronal layers interpenetrate each other. The interaction between the particles was quantified in terms of the sticky potential and showed a trend to be influenced by the particle size rather than the graft density of DNA strands on the particle.

Purpose: The aim of this research is to examine whether dental trainees’ stress-coping ability and health-maintenance capability affect their mental health during training. Methods: Trainee dentists who began their postgraduate clinical training for one year in 2010 (n = 60) and 2011 (n = 64) at Kyushu University Hospital were eligible to participate in the study. The questionnaire survey was conducted five times throughout the training year using the 13-item Sense of Coherence (SOC-13), the 28-item General Health Questionnaire (GHQ-28), and the 22-item Job Content Questionnaire (JCQ-22). Results: A 107 trainee dentists were completed (female n = 48, male n = 59). We identified the influence of SOC on the mental health status of postgraduate trainee dentists; that is, trainee dentists with high SOC showed favorable mental health, whereas those with low SOC showed poor mental health during the training. Concerning work conditions, trainee dentists with high SOC had adequate support from their supervisors and coworkers. Conclusion: This study showed the possibility of applying SOC for predicting the mental health status of trainee dentists. The results also suggest the need to introduce a solid support system from the early stages of training, in order to achieve the effects of mental health support for trainee dentists with low SOC.

A diblock copolymer containing a single-stranded DNA and a temperature-responsive poly(N-isopropylacrylamide) (PNIPAAm-b-DNA) was synthesized. The PNIPAAm segment was obtained with a low polydispersity by reversible addition–fragmentation chain transfer polymerization. The number-averaged molecular weight and the molecular weight distribution of the PNIPAAm were determined with size exclusion chromatography. PNIPAAm-b-DNA was prepared through the Michael addition reaction of thiol-terminated PNIPAAm to 5′-maleimide-modified DNA. The lower critical solution temperature (LCST) of the copolymer was determined with differential scanning calorimetry. PNIPAAm-b-DNA self-assembled above the LCST into the formation of a polymer micelle with a hydrophobic PNIPAAm core surrounded by a hydrophilic DNA corona. Both light scattering and transmittance measurements revealed that the colloidal stability of the single-stranded DNA-carrying polymer micelles was enormously decreased when the complementary DNA was added into the micelle’s dispersion to form the fully matched DNA duplex on the surface. It was also found that the colloidal stability of the micelle was extremely enhanced when the 5′-terminal-mismatched DNA was added to form a single-base mismatch at the interface between the DNA corona and the dispersal medium. The higher flexibility of the DNA terminus in the corona presumably increased the entropic repulsion between the micelles.

Diblock and miktoarm star-shaped thermoresponsive copolymers composed of single-stranded DNA (ssDNA) and poly(N-isopropylacrylamide) were successfully synthesized with combination of atom transfer radical polymerization (ATRP) and click chemistry. This approach should be generalizable to other DNA-functionalized copolymers. Such copolymers self-assemble into spherical micelles with ssDNA corona in aqueous solution above the lower critical solution temperature. The micellar size can be tuned from the macromolecular architecture. These DNA-encoded micellar particles are able to encapsulate and release hydrophobic guest molecules upon changing temperature.

A technique for tuning interparticle distance in plasmonic gold nanoparticle (AuNP) assemblies has shown great potential for the development of optoelectronic nanodevices. However, it still remains a challenge to reversibly alter the distance in a facile manner. DNA-templated AuNP assemblies are among the mostly investigated plasmonic nanomaterials. In previous work, salt-induced structural shrinkage of DNA-templated AuNP (5 nm in diameter) dimers/trimers is demonstrated only by electron microscopic analyses. In the present study, interparticle distance is modulated in larger AuNP (15 nm or 20 nm in diameter) dimers that exhibit strong surface plasmon resonance (SPR). The reversible SPR shift is achieved by using the temperature-dependent shrinkage/extension of the DNA-templated AuNP dimers. The present proof-of-concept study suggests a potential application of the reversible structural change to optical switching.

The shape of cells is a key determinant of cellular fates and activities. In this study, we demonstrate a method for controlling the cellular shape on a chemically modified glass coverslip with micropatterned cell adhesiveness. The glass surface was chemically modified with an alkylsiloxane monolayer having a caged carboxyl group, where single-cell-sized hydrophilic islands with hydrophobic background were created by irradiating the substrate in contact with a photomask to produce the carboxyl group. Thus, the created surface hydrophilicity pattern was converted to a negative pattern of a protein-repellent amphiphilic polymer, Pluronic F108, according to its preferential adsorption to the hydrophobic surfaces. The following addition of a cell-adhesive protein, fibronectin, resulted in its selective adsorption to the irradiated regions. In this way, cell-adhesive islands were produced reproductively, and the cells formed a given shape on the islands. As examples of the cell-shape control, we seeded HeLa ce...

Capillary electrophoretic separation of chemically synthesized ssDNA and a single-base-substituted one (normal and mutant ssDNA, respectively) was demonstrated using a PEG-oligodeoxyribonucleotide block copolymer (PEG-b-ODN) as an affinity ligand. When the base sequence of PEG-b-ODN was designed to be complementary to part of normal ssDNA including the base-substituted site, the electrophoretic mobility of normal ssDNA significantly decreased whereas that of mutant ssDNA slightly changed. Resolution of the separation strongly depended on the ODN length of the copolymer, the capillary temperature, and the Mg2+ concentration in the running buffer, indicating that the retardation of migration of normal ssDNA was induced by the reversible hybridization with PEG-b-ODN. It was found that the dissociation constant (K(d)) of the duplex between normal ssDNA and the affinity probe ODN should be smaller than 10(-6) M to achieve the good peak separation. In addition, we calculated the mobility of the complex (mu(C)) between normal ssDNA and PEG-b-ODN using a two-state model. The base sequence of affinity probe ODN appropriate to achieve the sufficient resolution will be predicted on the basis of the mu(C) and K(d )values.

Cell micropatterning is an important technique for a wide range of applications, such as tissue engineering, cell-based drug screening, and fundamental cell biology studies. This paper overviews cell patterning techniques based on chemically modified substrates with different degrees of cell adhesiveness. In particular, the focus is on dynamic substrates that change their cell adhesiveness in response to external stimuli, such as heat, voltage, and light. Such substrates allow researchers to achieve an in situ alteration of patterns of cell adhesiveness, which is useful for co-culturing multiple cell types and analyzing dynamic cellular activities. As an example of dynamic substrates, we introduce a dynamic substrate based on a caged compound, where we accomplished a light-driven alteration of cell adhesiveness and the analysis of a single cell's motility.

Purpose: γ-Aminobutyric acid (GABA) and N-methyl-D-aspartate (NMDA) receptors are important targets for anesthetic action at thein vitro cellular level. Gabaculine is a GABA-transaminase inhibitor that increases endogenous GABA in the brain, and enhances GABA activity. We have recently shown that unconsciousness is associated with the enhanced GABA activity due to gabaculine, but that immobility is not. MK-801 is a selective NMDA channel blocker. In this study, we examined behaviourally whether gabaculine in combination with MK-801 could produce these components of the general anesthetic state. We further compared the effect of MK-801 with ketamine, another NMDA channel blocker. Methods: All drugs were administered intraperitoneally to adult male ddY mice. To assess the general anesthetic components, two endpoints were used. One was loss of the righting reflex (LORR; as a measure of unconsciousness) and the other was loss of movement in response to tail-clamp stimulation (as a measure of immobility). Results: Large doses of MK-801 alone (10–50 mg·kg−1) induced neither LORR nor immobility in response to noxious stimulation. However, even a small dose (0.2 mg·kg−1) significantly enhanced gabaculine-induced LORR (P

An affinity capillary electrophoretic method was developed to detect a single-base difference of single-stranded DNA (ssDNA). Poly(ethylene glycol)-oligodeoxyribonucleotide block copolymers (PEG-b-ODN) were prepared for use as a novel affinity ligand. We introduced a running buffer solution of PEG-b-ODN into a capillary tube, and electrophoretically separated a mixture of chemically synthesized 20 mer ssDNA (normal ssDNA) and a single-base-substituted 20 mer ssDNA (mutant ssDNA). When the base sequence of PEG-b-ODN was designed to be complementary to part of the normal ssDNA, the migration rate of the normal ssDNA was significantly decreased by reversible hybridization with PEG-b-ODN, depending on the base number of PEG-b-ODN, the salt concentration of the running buffer, and the capillary temperature. In contrast, the mobility of mutant ssDNA did not change because the interaction with PEG-b-ODN was negligible. Optimization of the analytical conditions gave two distinct peaks, one for normal and the other for mutant ssDNA, on the electropherogram, allowing for facile discrimination of the single-base difference. The results indicate that PEG-b-ODN is a promising affinity ligand for the capillary electrophoretic separation of normal and single-base mutated ssDNA.

Monitoring and assessing of patient respiratory function during conscious sedation are important because many drugs used for conscious sedation produce respiratory depression and subsequent hypoventilation. The purpose of this study is to assess the value of a dynamic air-pressure sensor for respiratory monitoring of clothed patients. Eight clothed adult volunteers were reclined on a dental chair positioned horizontally. The air bag for measuring air-pressure signals corresponding to respiration was placed on the seat back of the dental chair in the central lumbar area of the subject. The subject breathed through a face mask with a respirometer attached for measuring expiratory tidal volume. The air-pressure signals corresponding to respiration were obtained and the time integration values for air pressure during each expiration (integral P(exp)) were calculated. The expiratory tidal volume (TV(exp)) was measured simultaneously by respirometer. The relationship between TV(exp) and integral P(exp) for each subject was assessed by a Pearson correlation coefficient. A strong correlation between TV(exp) and integral P(exp) was observed in all subjects. Measuring integral P(exp) by dynamic air-pressure sensor makes it possible to estimate respiratory volume breath by breath, and the respiratory pressure-time integral waveform was useful in visually monitoring the respiration pattern. We believe that in the future this device will be used to monitor respiratory physiology in clothed patients, contributing to safer sedative procedures.

Gold nanoparticles modified with DNA duplexes are rapidly and spontaneously aggregated at high ionic strength. In contrast, this aggregation is greatly suppressed when the DNA duplex has a single-base mismatch or a single-nucleotide overhang located at the outermost surface of the particle. These colloidal features emerge irrespective of the size and composition of the particle core; however, the effects of the shape remain unexplored. Using gold nanorods and nanotriangles (nanoplatelets), we show herein that both remarkable rapidity in colloidal aggregation and extreme susceptibility to DNA structural perturbations are preserved, regardless of the shape and aspect ratio of the core. It is also demonstrated that the DNA-modified gold nanorods and nanotriangles are applicable to naked-eye detection of a single-base difference in a gene model. The current study corroborates the generality of the unique colloidal properties of DNA-functionalized nanoparticles, and thus enhances the feasibility of their practical use.

The beta(2) adrenergic receptor (beta(2)AR) is a G protein-coupled receptor that is selective to epinephrine. We demonstrate herein monitoring of an agonist-induced conformational change of beta(2)AR in living cells. The monitoring method is based on fluorescence resonance energy transfer from a cyan fluorescent protein (CFP) to a biarsenical fluorophore, FlAsH, attached to the C-terminus, and the third intracellular loop (ICL3), respectively. Recombinant beta(2)ARs exhibited agonist-induced increases in the FlAsH/CFP emission ratio, indicating that the ICL3 approached the C-terminus upon activation. Since the emission ratio changes were on a time scale of seconds, the conformational change of beta(2)AR in living cells was more rapid than that of purified beta(2)AR measured in vitro. Interestingly, the direction of the emission ratio change of beta(2)AR was opposite to that of the norepinephrine-responsive alpha(2A) adrenergic receptor reported recently. It was suggested that this discrepancy corresponds directly to the diametric biological functions, i.e., the activation or inactivation of adenylyl cyclase.

We developed a weak-affinity separation system for single-nucleotide polymorphisms (SNPs) based on capillary electrophoresis. In this approach, single-stranded DNA (ssDNA)-polyacrylamide (polyAAm) conjugate was used as a pseudo-immobilized affinity ligand to separate the target DNA, cytochrome P450 2C9 (CYP2C9), and its point mutant. The ligand DNA was designed to be complementary to the normal DNA, and the target DNA was electrophoretically separated by the difference in the affinity with the pseudo-immobilized ligand in the capillary. We showed that the separation efficiency was closely associated with the Tm value of double-stranded DNA (dsDNA) consisting of the target and ligand DNA, which depends on the measurement conditions, such as the base number of the ligand DNA and the concentration of Mg2+ in the buffer solution.