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Here we report a bio-adhesive porous organic module (Glue COF) composed of hexagonally packed 1D nanopores based on a covalent organic framework. The nanopores are densely decorated with guanidinium ion (Gu+ ) pendants capable of forming salt bridges with oxyanionic species. Glue COF strongly adheres to biopolymers through multivalent salt-bridging interactions with their ubiquitous oxyanionic species. By taking advantage of its strong bio-adhesive nature, we succeeded in creating a gate that possibly opens the nanopores through a selective interaction with a reporter chemical and releases guest molecules. We chose calmodulin (CaM) as a gating component that can stably entrap a loaded guest, sulforhodamine B (SRB), within the nanopores (CaM COF⊃SRB). CaM is known to change its conformation on binding with Ca2+ ions. We confirmed that mixing CaM COF⊃SRB with Ca2+ resulted in the release of SRB from the nanopores, whereas the use of weakly binding Mg2+ ions resulted in a much slower release of SRB.

Anion-π interaction has been found to play a key role in interfacial phenomena. In this study, we evaluated the anion-π interactions at the solid/water interface. Anion adsorption originating from anion-π interaction at the interfaces followed the hydration energy and the presence of conjugated systems of the anions by the QCM measurements. Force curve measurements revealed that the single-molecule force of anion-π interaction between an NDI unit and the negatively charged surface of the cantilever was ∼40 pN. To the best of our knowledge, this is the first example of obtaining a single-molecule force for anion-π interactions.

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Herein we report the synthesis and isolation of a shape-persistent Janus protein nanoparticle derived from the biomolecular machine chaperonin GroEL (^AGroEL^B) and its application to DNA-mediated ternary supramolecular copolymerization. To synthesize ^AGroEL^B with two different DNA strands A and B at its opposite apical domains, we utilized the unique biological property of GroEL, i.e., Mg²⁺/ATP-mediated ring exchange between ^AGroEL^A and ^BGroEL^B with their hollow cylindrical double-decker architectures. This exchange event was reported more than 24 years ago but has never been utilized for molecular engineering of GroEL. We leveraged DNA nanotechnology to purely isolate Janus ^AGroEL^B and succeeded in its precision ternary supramolecular copolymerization with two DNA comonomers, A** and B*, that are partially complementary to A and B in ^AGroEL^B, respectively, and programmed to self-dimerize on the other side. Transmission electron microscopy allowed us to confirm the formation of the expected dual-periodic copolymer sequence −(^(B*/B)GroEL^(A/A**/A**/A)GroEL^(B/B*))– in the form of a laterally connected lamellar assembly rather than a single-chain copolymer.

Objective: The objective of this study was to clarify the learning effect of a tooth brushing practice (TBP) module resembling the oral environment developed to easily practice brushing of patients’ teeth.

A speciation method for chromate, selenite, and selenate was developed by high-performance liquid chromatography (HPLC)–fluorometric detection (FL) using a single derivatizing reagent. Chromium(VI)...

This course presents an elementary, intuitive, and visual introduction to several mathematical basics for beginners in computer graphics. The mathematical concepts covered in this course include 2D/3D translation, rotation, affine transformation, quaternion, dual quaternion, exponential and logarithm. These are quite useful for various aspects of computer graphics, including curve/surface editing, deformation and animation of geometric objects. On one hand we don't assume that the audience has familiarity with such mathematical concepts. Elements of linear algebra and calculus at undergraduate level are sufficient for the attendees of this course. To clarify the relation between these concepts and computer graphics practice, several graphics techniques will also be demonstrated, such as morphing (ARAP), cage-based deformation and Poisson mesh editing.

The scavenging effects of metal complexes of thiacalix[4]arenetetrasulfonate (Me-TCAS[4], Me=H₂, Fe³(+), Mn³(+), Mn²(+), Cu²(+), and Zn²(+)) on superoxide anion radicals (O₂⁻) generated from the xanthine-xanthine oxidase system were investigated by the nitroblue tetrazolium (NBT) method and electron spin resonance (ESR) spin-trapping method using 5,5-dimethyl-1-pyrroline-N-oxide as a trapping reagent. As a reference, calix[4]arenetetrasulfonate (H₂)-CAS[4]), calix[6]arenehexasulfonate (H₂-CAS[6]) and calix[8]areneoctasulfonate (H₂-CAS[8]) were also examined. The results by the NBT method indicated that Fe³(+)- and Mn³(+)-TCAS[4] exhibited the highest O₂⁻ scavenging activity among Me-TCAS[4] and H₂-CAS[n] (n = 4, 6, 8) in this study. The IC₅₀ values of Fe³(+)- and Mn³(+)-TCAS[4] for O₂⁻ scavenging activity were estimated to be 5.3 and 7.8 µM, respectively, and were almost the same as those of tannin acid, catechin and their derivatives, which are known as very effective scavengers of O₂⁻. Scavenging activities were in the order of Fe³(+)- and Mn³(+)-TCAS[4]>>Mn²(+)-, Cu²(+)-, and Zn(2+)-TCAS[4]>>H(2)-TCAS[4] and H₂-CAS[n] (n=4, 6, 8). Each activity of Me-TCAS[4] (Me=Fe³(+), Mn³(+), Mn²(+), Cu²(+), and Zn²(+)) was higher than that of the corresponding metal ion, indicating that H₂-TCAS[4] has the ability to raise the activity of the metal ion itself by forming a complex. Also, the ESR spin-trapping method revealed that Fe³(+)- and Mn³(+)-TCAS[4] showed high O₂⁻ scavenging activities, similarly to the results by the NBT method.

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.

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.

We researched the developmental problems on a case of deaf child with autism, and compared his development with a case of deaf-single child. We followed the cases through the training in our clinic since age of 3 to the age of 6, and assessed them by the developmental test of Enjhoji, and Tanaka-Binet intelligence Scale, and we also assessed them by WISC-III for reference at finally. In the case-B; the deaf-single child, he could catch up with the normal level on all developmental abilities assessed by these tests. In the case-A; the deaf child with autism, it was remarkable that the retardation of the abilities of transferable-movement, the emotional development, the verbal-intelligence. It was suggested that such retardations of case-A were mainly caused by the autism, in spite of he had started wearing hearing-aids from age of 1.

Virulent Rhodococcus equi is an important pathogen in foals aged 1 to 3 months, but its incidence in Tennessee, U. S. A., is poorly understood. Ten soil samples were collected from a Tennessee walking horse farm in Tennessee. The presence of R. equi was investigated and 107 isolates were tested for the presence of 15- to 17-kDa antigens (VapA) by immunoblotting and PCR. R. equi was isolated (103-104 colony forming units/g) from 8 of the 10 soil samples. Twenty-four VapA-positive isolates were detected from 7 of the 8 culture-positive samples with a prevalences ranging from 6.7 to 50%. Of the 24 virulent isolates, 21 contained an 85-kb type I plasmid, and 3 contained an 85-kb type III plasmid which had been uniquely isolated from soil isolates in Texas. There was no evidence of R. equi infection on the farm, but the virulent form was widespread in the soil environment of the farm.

We investigated the prevalence of virulent Rhodococcus equi in clinical isolates from 69 sporadic cases (60 men, 8 women, and 1 patient of unknown sex) in Chiang Mai, Thailand, between 1993 and 2001. Fifty were human immunodeficiency virus (HIV) positive, 3 were HIV negative, and HIV status was unknown for 16. Fifty-two (75%) of 69 isolates were strains of intermediate virulence that contained the virulence-associated 20-kDa antigen, and 17 isolates (25%) were avirulent. No virulent strains with the virulence-associated 15-17-kDa antigens were identified. R. equi was isolated from HIV-positive patients' houses and those of their neighbors: avirulent strains were widespread, but only 1 strain of intermediate virulence was isolated. R. equi strains of intermediate virulence were isolated from 4 (0.8%) of 500 submaxillary lymph nodes from apparently healthy pigs in Chiang Mai. The routes of R. equi acquisition should be investigated from the viewpoint of zoonosis and public health.

Diblock copolymers composed of allele-specific oligodeoxyribonucleotide (ODN) and poly(ethylene glycol) (PEG) are used as an affinity probe of free-solution capillary electrophoresis to quantitatively detect single-base substitutions in genetic samples. During electrophoresis, the probe binds strongly to a wild-type single-stranded DNA analyte (WT) through hybridization, while it binds weakly to its single-base-mutated DNA analyte (MT) due to a mismatch. Complex formation with the probe augments the hydrodynamic friction of either analyte, thereby retarding its migration. The difference in affinity strength leads to separation of the WT, MT, and contaminants, including the PCR primers used for sample preparation. The optimal sequence of the probe's ODN segment is rationally determined in such a way that the binding constant between the ODN segment and MT at the capillary temperature is on the order of 10(6) M(-1). The validity of this guideline is verified using various chemically synthesized DNA analytes, as well as those derived from a bacterial genome. The peak area ratio of MT agrees well with its feed ratio, suggesting the prospective use of the present method in SNP allele frequency estimation.

Capillary electrophoretic separation of a mixture of 5'-fluorescein isothiocyanate-labeled single-stranded DNA (normal ssDNA) and its single-base-substituted one (mutant ssDNA) was achieved by using a RAFT-generated polyacrylamide-oligodeoxyribonucleotide block copolymer (PAAm-b-ODN) as an affinity polymeric probe. PAAm-b-ODN was synthesized through the Michael addition of thiol-terminated PAAm (PAAm-SH) to 5'-maleimide-modified ODN. PAAm-SH was derived from dithiobenzoate-terminated PAAm prepared via RAFT polymerization. The number-averaged molecular weight (M(n)) and the molecular weight distribution were determined by aqueous size exclusion chromatography. After a capillary tube was filled with the running buffer solution of PAAm-b-ODN, a mixture of normal and mutant ssDNA was subjected to electrophoresis and detected by a laser-induced fluorescent detector. Because the base sequence of PAAm-b-ODN was complementary to part of the mutant ssDNA, including a single-base substitution site, the electrophoretic migration of mutant ssDNA was retarded due to the formation of the equilibrium complex with PAAm-b-ODN. Increasing M(n) of the PAAm segment enhanced this retardation. On the other hand, normal ssDNA was unable to form the complex owing to a single-base mismatch, which was proved by melting curve measurements. The Lineweaver-Burk-type analysis of the mobility of mutant ssDNA revealed that the binding constants for the complexes with different PAAm-b-ODN probes were almost identical to each other. The analysis also demonstrated that the ratio of the hydrodynamic radius of the complex to that of the free mutant ssDNA increased with increasing M(n) of the affinity polymeric probe's PAAm segment. This means that the PAAm segment indirectly provides mutant ssDNA with an additional hydrodynamic friction force via the affinity interaction of the ODN segment. Optimization of the salt concentration of the running buffer and the capillary temperature improved the resolution of the separation. This affinity polymeric probe will be useful for developing a simple and highly reliable single-nucleotide polymorphism genotyping method.

Capillary electrophoretic separation of 60 mer single-stranded DNA (ssDNA) and a single-base-substituted ssDNA was demonstrated using a size- and composition-controlled poly(ethylene glycol)-oligodeoxyribonucleotide block copolymer (PEG-b-ODN) as an affinity ligand. Under appropriate conditions, PEG-b-ODN and ssDNA with a complementary sequence formed a reversible complex via hybridization during the electrophoresis, while the copolymer did not interact with the single-base-substituted ssDNA. The copolymer's PEG length determined the electrophoretic mobility of the ssDNA; upon formation of the complex, the electrically neutral PEG added hydrodynamic friction to ssDNA. Simultaneously using two types of PEG-b-ODN copolymers whose PEG segments were of different lengths, we achieved the complete separation of the 60 mer ssDNA, its single-base-substituted ssDNA, and impurities. This method was sensitive enough to quantify a slight amount (approximately 1%) of the single-base-substituted ssDNA. The present results suggest that our approach is applicable to quantitative detection of minor genotypes.

application/pdf, Influenza viruses infect vertebrates, including mammals and birds. Influenza virus reverse-genetics systems facilitate the study of the structure and function of viral factors. In contrast, less is known about host factors involved in the replication process. Here, we developed a replication and transcription system of the negativestrand RNA genome of the influenza virus in Saccharomyces cerevisiae, which depends on viral RNAs, viral RNA polymerases, and nucleoprotein (NP). Disruption of SUB2 encoding an orthologue of human RAF-2p48/UAP56, a previously identified viral RNA synthesis stimulatory host factor, resulted in reduction of the viral RNA synthesis rate. Using a genome-wide set of yeast single-gene deletion strains, we found several host factor candidates affecting viral RNA synthesis. We found that among them, Tat-SF1, a mammalian homologue of yeast CUS2, was a stimulatory host factor in influenza virus RNA synthesis. Tat-SF1 interacted with free NP, but not with NP associated with RNA, and facilitated formation of RNA-NP complexes. These results suggest that Tat-SF1 may function as a molecular chaperone for NP, as does RAF-2p48/UAP56. This system has proven useful for further studies on the mechanism of influenza virus genome replication and transcription.

PURPOSE To compare visual acuity measures obtained with the M&S Technologies Smart System II (SSII) and the revised Early Treatment of Diabetic Retinopathy Study (ETDRS) charts in terms of accuracy and test-retest repeatability. METHODS Monocular visual acuities were taken in 57 young, visually normal adults on two separate visits in which both the SSII system and the ETDRS charts were tested in random order by two masked examiners. The eye to be tested throughout was chosen randomly at the initial visit. Measurements were made through an optimal phoropter correction, determined by a noncycloplegic refraction for a 10-foot distance. Both charts were presented at 10 feet, and were matched closely for luminance. RESULTS The mean visual acuity in the group was -0.16 log minimum angle of resolution (MAR) for the ETDRS chart and -0.18 log MAR for the SSII, a small but statistically significant difference. A 95% confidence interval for the mean difference in visual acuity between the two charts was -0.033 log MAR to -0.003 log MAR. The test-retest repeatability was not significantly different in the two tests. The 95% limits of agreement for test-retest repeatability were -0.13 log MAR to +0.17 log MAR for the SSII and -0.12 log MAR to +0.13 log MAR for the ETDRS charts. CONCLUSIONS The SSII can provide an accurate (mean difference

Rhodococcus equi was isolated from fecal and soil samples from four native Jeju horse farms and six Thoroughbred farms in Jeju, Korea. The isolates were examined for the presence of virulence-associated 15-17-kDa antigens (VapA) by colony blotting, using the monoclonal antibody 10G5, and for the gene encoding VapA by PCR. R. equi was isolated from all 36 soil samples collected from the 10 farms with between 5.0 x 10(2) and 7.5 x 10(4) colony-forming units (cfu) per gram of soil, and from 37 of 40 fecal samples with between 5.0 x 10(1) and 1.1 x 10 (5) cfu per gram of feces. Virulent R. equi was isolated from seven farms and appeared in 2.0% of isolates (10 of 508). Of the 10 virulent isolates, four contained a 90-kb type II plasmid, which has been found in isolates from the Kiso native horses of Japan, and the other six contained a new variant, which did not display the EcoRI and EcoT22I digestion patterns of the 10 representative plasmids already reported (85-kb types I, II, III, and IV; 87-kb types I and II; 90-kb types I, II, III, and IV). We designated the new variant as the "90-kb type V" plasmid, because its EcoRI digestion pattern is similar to that of the 90-kb type II plasmid. This is the first report of the prevalence of virulent R. equi in Jeju, Korea. The same virulence plasmid type is found in both Korean and Japanese isolates, providing insight into the origin, ancestry, and dispersal of native horses in Korea and Japan.

An affinity capillary electrophoresis was developed for detecting a point mutation of single-stranded DNA (ssDNA). Poly(ethylene glycol)-oligodeoxyribonucleo-tide block copolymers (PEG-b-ODN) were prepared to use as a novel affinity ligand. Optimization of the analytical conditions, such as the salt concentration of running buffer, the capillary temperature and the base number of the copolymer, gave two distinct peaks for wild-type (WT) and point-mutated ssDNA (MT) on the electropherogram, allowing for the facile discrimination of the single-base difference.

The presentation deals with the calibration for a biological acoustic impedance microscope. A block of tissue was placed on a plastic substrate, and an acoustic pulse focused on the interface was transmitted from the rear side of the substrate. The reflection was received by the same transducer and interpreted into local acoustic impedance. An acoustic impedance image was obtained by mechanically scanning the transducer. A reference material was simultaneously observed in order to perform calibration. A water droplet was employed as a reference material, however, it often entered into the interface between the substrate and tissue, leading to the reduction of the image quality. Therefore, several kinds of compounds were investigated. Silicone rubber was relatively homogeneous and stable in acoustic impedance, after a certain period of curing time. However, its dispersion with the scale of focal spot size was as large as 0.02 Ns/cm3, suggesting that a sufficient number of points should be measured and aver...