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Structures of monothiol and dithiol glutaredoxin-1 from C. oremlandii are reported. Monothiol glutaredoxin-1 formed domain-swapped structures, one of which was in a hetero-oligomer complex., Protein dimerization or oligomerization resulting from swapping part of the protein between neighboring polypeptide chains is known to play a key role in the regulation of protein function and in the formation of protein aggregates. Glutaredoxin-1 from Clostridium oremlandii (cGrx1) was used as a model to explore the formation of multiple domain-swapped conformations, which were made possible by modulating several hinge-loop residues that can form a pivot for domain swapping. Specifically, two alternative domain-swapped structures were generated and analyzed using nuclear magnetic resonance (NMR), X-ray crystallography, circular-dichroism spectroscopy and hydrogen/deuterium-exchange (HDX) mass spectrometry. The first domain-swapped structure (β3-swap) was formed by the hexameric cGrx1–cMsrA complex. The second domain-swapped structure (β1-swap) was formed by monothiol cGrx1 (C16S) alone. In summary, the first domain-swapped structure of an oxidoreductase in a hetero-oligomeric complex is presented. In particular, a single point mutation of a key cysteine residue to serine led to the formation of an intramolecular disulfide bond, as opposed to an intermolecular disulfide bond, and resulted in modulation of the underlying free-energy landscape of protein oligomerization.

This paper investigates the correlation between the random lasing properties of organic waveguides made by poly-(9-9dioctylfluorene) (PFO) thin films and the morphology of wrinkled corrugated substrates. The capability to individually control the wrinkle wavelength, shape, and height allows us to separately investigate their role on the sample emission properties. We demonstrate that the main parameter determining the presence of coherent random lasing is the substrate roughness and that, contrary to what could be qualitatively expected, as the roughness increases, coherent random lasing is progressively reduced. Coherent random lasing is observed only for a substrate roughness below 33 nm, while higher roughness leads to amplified spontaneous emission (up to 70 nm) or to the absence of light amplification in the film (above 70 nm). We demonstrate that this result is due to a progressive reduction of the light amplification efficiency in the PFO film, evidencing that coherent random lasing can be obtained only with a right interplay between light amplification and scattering. Besides clarifying the basic aspects of random lasing in organic waveguides, our work opens the way to the realization of organic random lasers with predictable emission properties, thanks to the high control level of the scattering properties of the wrinkled corrugated surfaces.

We investigated strontium oxide (SrO) as an interlayer material to enhance the UV stability of a CH3NH3PbI3 perovskite solar cell. Moisture and over 400 nm wavelength of light were excluded to investigate the effect of UV light only. Two different interlayer fabrication processes were examined to optimize the performance of this solar cell. Devices fabricated by dipping for 30 min in SrO solution exhibited photoconversion efficiencies of 15.5%, whereas those fabricated with 60-min dipping showed photoconversion efficiencies of 15% and exhibited local Sr agglomeration. Devices with SrO displayed lower initial efficiencies than those without any SrO layer (17.6%), However, a device without SrO retained only 34.4% of its initial efficiency after 100 h of UV exposure. In contrast, SrO-incorporated devices retained almost 60.0% of their initial efficiency. Severe μ-PL mapping intensity degradation was observed in devices that did not include the interlayer, but no degradation was observed in those with the SrO interlayer. This can be attributed to the passivation of the degradation sites by SrO.

The effects of tyrosine kinase inhibitors (TKIs) were evaluated on growth inhibition of intracellular Toxoplasma gondii in host ARPE-19 cells. The number of tachyzoites per parasitophorous vacuolar membrane (PVM) was counted after treatment with TKIs. T. gondii protein expression was assessed by western blot. Immunofluorescence assay was performed using Programmed Cell Death 4 (PDCD4) and T. gondii GRA3 antibodies. The TKIs were divided into 3 groups; non-epidermal growth factor receptor (non-EGFR), anti-human EGFR 2 (anti-HER2), and anti-HER2/4 TKIs, respectively. Group I TKIs (nintedanib, AZD9291, and sunitinib) were unable to inhibit proliferation without destroying host cells. Group II TKIs (lapatinib, gefitinib, erlotinib, and AG1478) inhibited proliferation up to 98% equivalent to control pyrimethamine (5 μM) at 20 μM and higher, without affecting host cells. Group III TKIs (neratinib, dacomitinib, afatinib, and pelitinib) inhibited proliferation up to 98% equivalent to pyrimethamine at 1-5 μM, but host cells were destroyed at 10-20 μM. In Group I, TgHSP90 and SAG1 inhibitions were weak, and GRA3 expression was moderately inhibited. In Group II, TgHSP90 and SAG1 expressions seemed to be slightly enhanced, while GRA3 showed none to mild inhibition; however, AG1478 inhibited all proteins moderately. Protein expression was blocked in Group III, comparable to pyrimethamine. PDCD4 and GRA3 were well localized inside the nuclei in Group I, mildly disrupted in Group II, and were completely disrupted in Group III. This study suggests the possibility of a vital T. gondii TK having potential HER2/4 properties, thus anti-HER2/4 TKIs may inhibit intracellular parasite proliferation with minimal adverse effects on host cells.

We characterize wrinkling on the surfaces of prestrained polystyrene sheets coated with thin polytetrafluoroethylene skins using a combination of mechanical strain measurements and 3D finite element simulations. The simulations show that wrinkle wavelength increases with skin thickness, in agreement with a well-known continuum model and recent experiments. The wrinkle amplitudes also increase with strain. Nanoinclusions, such as holes and patterned lines, influence wrinkle patterns over limited distances, and these distances are shown to scale with the wrinkle wavelengths. Good agreement between experimental and simulated influence distances is observed. The inclusions provide strain relief, and they behave as if they are attracting adjacent material when the sheets are under strain. The wrinkles have stiffnesses in much the same way as do polymers (but at different length scales), a property that is quantified for polymers using persistence lengths. We show that the concept of persistence length can be useful in characterizing the wrinkle properties that we have observed. However, the calculated persistence lengths do not vary systematically with thickness and strain, as interactions between neighboring wrinkles produce confinement that is analogous to the kinetic confinement of polymers.

We show that submicron-sized patterns can be imprinted into soft, recombinant-engineered protein hydrogels (here elastin-like proteins, ELP) by transferring wavy patterns from polydimethylsiloxane (PDMS) molds. The high-precision topographical tunability of the relatively stiff PDMS is translated to a bio-responsive, soft material, enabling topographical cell response studies at elastic moduli matching those of tissues. Aligned and unaligned wavy patterns with mold periodicities of 0.24-4.54 μm were imprinted and characterized by coherent anti-Stokes Raman scattering and atomic force microscopy. The pattern was successfully transferred down to 0.37 μm periodicity (width in ELP: 250 ± 50 nm, height: 70 ± 40 nm). The limit was set by inherent protein assemblies (diameter: 124-180 nm) that formed due to lower critical solution temperature behavior of the ELP during molding. The width/height of the ELP ridges depended on the degree of hydration; from complete dehydration to full hydration, ELP ridge width ranged from 79 ± 9% to 150 ± 40% of the mold width. The surface of the ridged ELP featured densely packed protein aggregates that were larger in size than those observed in bulk/flat ELP. Adipose-derived stem cells (ADSCs) oriented along hydrated aligned patterns with periodicities ≥0.60 μm (height ≥170 ± 100 nm), while random orientation was observed for smaller distances/amplitudes, as well as flat and unaligned wavy ELP surfaces. Hence, micro-molding of ELP is a promising approach to create tissue-mimicking, hierarchical architectures composed of tunable micron-sized structures with nano-sized protein aggregates, which opens the way for orthogonal screening of cell responses to topography and cell-adhesion ligands at relevant elastic moduli.

Human lysophosphatidic acid receptor 1 (LPA1 ) is a G-protein coupled receptor that mediates various biological functions such as proliferation, platelet aggregation, smooth muscle contraction, and tumor cell invasion. For dissection of the molecular function of LPA1 , a recombinant LPA1 was overexpressed in Escherichia coli membrane fractions and purified to homogeneity by single affinity chromatography. The purified LPA1 was stabilized with an amphiphilic polymer that was synthesized by the coupling of octylamine, glucosamine, and diethylaminoproylamine at the carboxylic groups of poly-γ-glutamic acid. The complex of purified LPA1 and amphiphilic polymer showed a monodisperse oligomer and specific binding to LPA with apparent Ki values of 30 μM. Compared with the Gs protein, it also showed selective binding to the alpha subunit of the Gi protein. These results indicate that recombinant LPA1 in an amphiphilic polymer complex has an active conformation for interaction with ligands and G-proteins.

This paper reports a scalable approach to achieve spatially selective graphene functionalization using multiscale wrinkles. Graphene wrinkles were formed by relieving the strain in thermoplastic polystyrene substrates conformally coated with fluoropolymer and graphene skin layers. Chemical reactivity of a fluorination process could be tuned by changing the local curvature of the graphene nanostructures. Patterned areas of graphene nanowrinkles and crumples followed by a single-process plasma reaction resulted in substrates with regions having different fluorination levels. Notably, conductivity of the functionalized graphene nanostructures could be locally tuned as a function of feature size without affecting the mechanical properties.

Nanoscale wrinkling on the surfaces of polymer-based materials can be precisely controlled by depositing thin metal films of varying thicknesses. The deposition of these films fundamentally alters the mechanical properties of the substrates in ways that are not simply described using traditional continuum mechanical frameworks. In particular, we find, by modeling within a finite element analysis approach, that the very act of depositing a metal film may alter the Young's modulus of the polymer substrate to depths of up to a few hundred nanometers, creating a modified interfacial skin layer. We find that simulated wrinkle patterns reproduce the experimentally observed features only when the modulus of this surface layer varies by more than ∼500 nm and is described using a sigmoidal gradient multiplier.

Transglutaminase 2 (TGase 2) catalyzes a crosslink between protein bound-glutamine and -lysine. We proposed the mechanism of TGase 2 activation depends on conformation change from unfolded monomer to unfolded dimer. We found that TGase 2 has temperature-sensitive conformation change system at 30 °C. Small-angle X-ray scattering analysis showed that the enzyme was maintained as an unfolded monomer at temperatures below 30 °C, but changed to an unfolded dimer at over 30 °C. Mass analysis revealed that the C-terminus of TGase 2 was the critical region for dimerization. Furthermore, this conformational switch creates new biochemical reactivity that catalyzed inter-molecular crosslink at above 30 °C as an unfolded dimer of TGase 2 while catalyzed intra-molecular crosslink at below 30 °C as an unfolded monomer of TGase 2. The mechanism of TGase 2 activation depends on temperature-sensitive conformation change from unfolded monomer to unfolded dimer at over 30 °C. Furthermore, inter-molecular crosslinking activity is generated by the dimeric form of TGase 2. TGase 2 switches its conformation from a monomer to a dimer following a change in temperature, which engendered unique catalytic function of enzyme as inter-molecular crosslinking activity with calcium.

In general, expression of transglutaminase 2 (TGase 2) is upregulated in renal cell carcinoma (RCC), resulting in p53 instability. Previous studies show that TGase 2 binds to p53 and transports it to the autophagosome. Knockdown or inhibition of TGase 2 in RCC induces p53-mediated apoptosis. Here, we screened a chemical library for TGase 2 inhibitors and identified streptonigrin as a potential therapeutic compound for RCC. Surface plasmon resonance and mass spectroscopy were used to measure streptonigrin binding to TGase 2. Mass spectrometry analysis revealed that streptonigrin binds to the N-terminus of TGase 2 (amino acids 95&ndash, 116), which is associated with inhibition of TGase 2 activity in vitro and with p53 stabilization in RCC. The anti-cancer effects of streptonigrin on RCC cell lines were demonstrated in cell proliferation and cell death assays. In addition, a single dose of streptonigrin (0.2 mg/kg) showed marked anti-tumor effects in a preclinical RCC model by stabilizing p53. Inhibition of TGase 2 using streptonigrin increased p53 stability, which resulted in p53-mediated apoptosis of RCC. Thus, targeting TGase 2 may be a new therapeutic approach to RCC.

Soft skin layers on elastomeric substrates are demonstrated to support mechano-responsive wrinkle patterns that do not exhibit cracking under applied strain. Soft fluoropolymer skin layers on pre-strained poly(dimethylsiloxane) slabs achieved crack-free surface wrinkling at high strain regimes not possible by using conventional stiff skin layers. A side-by-side comparison between the soft and hard skin layers after multiple cycles of stretching and releasing revealed that the soft skin layer enabled dynamic control over wrinkle topography without cracks or delamination. We systematically characterized the evolution of wrinkle wavelength, amplitude, and orientation as a function of tensile strain to resolve the crack-free structural transformation. We demonstrated that wrinkled surfaces can guide water spreading along wrinkle orientation, and hence switchable, anisotropic wetting was realized.

Difficulties in the extraction of membrane proteins from cell membrane and their solubilization in native conformations have hindered their structural and biochemical analysis. To overcome these difficulties, an amphipathic polypeptide was synthesized by the conjugation of octyl and glucosyl groups to the carboxyl groups of poly-γ-glutamic acid (PGA). This polymer, called amphipathic PGA (APG), self-assembles as mono-disperse oligomers consisted of 4–5 monomers. APG shows significantly low value of critical micelle concentration and stabilization activity toward membrane proteins. Most of the sodium dodecyl sulfate (SDS)-solubilized membrane proteins from Escherichia coli remain soluble state in the presence of APG even after the removal of SDS. In addition, APG stabilizes purified 7 transmembrane proteins such as bacteriorhodopsin and human endothelin receptor Type A (ETA) in their active conformations. Furthermore, ETA in complex with APG is readily inserted into liposomes without disrupting the integrity of liposomes. These properties of APG can be applied to overcome the difficulties in the stabilization and reconstitution of membrane proteins.

Bacterial resistance to many existing antibiotics is a growing health concern worldwide. There is an urgent need to identify new antibiotics with unexploited modes of action. Peptide deformylase (PDF) is an essential enzyme involved in N-terminal protein processing in eubacteria but not in higher organisms. Therefore, PDF is considered an attractive target for the development of novel antibiotics. Here, we report the structures of the PDFs from Enterococcus faecalis (EfPDF) and Streptococcus pyogenes (SpyPDF) complexed with actinonin at 1.4 and 2.1 A resolutions, respectively. Actinonin, a naturally occurring, highly potent inhibitor, is bound tightly at the active site. The conformation of actinonin in the EfPDF and SpyPDF complexes was similar to those of all others. The detailed information from this study will facilitate the development of novel antibacterial molecules.

The main function of methionine sulfoxide reductases (Msr) in many organisms is to protect cells against oxidative stress caused by the catalyzed reduction of oxidized methionine to normal methionine. In a few micro-organisms, the existence of Msr as a fusion protein on a single polypeptide, MsrAB, was reported. However, Msr generally exists as separate enzymes MsrA and MsrB. Here, MsrAB from Helicobacter pylori (HpMsrAB) was overexpressed in Escherichia coli, purified, and crystallized to determine its structure. HpMsrAB X-ray diffraction data were collected to the resolution of 3.3 A, and the crystals were found to belong to the tetragonal space group P41212, with the unit cell parameters a =100.91, b =100.91, and c =160.08 A. The crystals corresponded to 5.38 A3 Da−1 of Matthews coefficient and 77.2% solvent content from the molecular replacement suggest that there is a single molecule in an asymmetric unit. Due to their unusually high solvent content, diffraction of these crystals only reach a resolution of 3.3 A. A preliminary solution was determined by molecular replacement. Further refinement of the structure is in progress.

Single-chain variable fragments of antibodies (scFv) specific to 2,4-dinitrotoluene (DNT) were isolated from a phage library displaying synthetic human scFv fragments with 6 diversified complementary determining regions (CDRs). A DNT derivative that contained an extended amine group was synthesized and conjugated to the NHS-group that was linked to magnetic beads. Phages specific to the immobilized DNT derivatives were isolated from the library after 4 rounds of sequential binding and elution processes. The displayed scFv fragments from the isolated phages showed consensus CDR sequences. One DNT-specific scFv was expressed in E. coli and purified using Ni-affinity chromatography. The purified DNT-specific scFv binds specifically to the immobilized DNT-derivative with value of M. The scFv and DNT interaction was not disrupted by the addition of 4-nitrotoluene or benzoic acid. These data demonstrate that the screened scFv from the phage displayed library could be used for selective and sensitive detection of explosives such as TNT.

Vertical alignment (VA) of a liquid crystal display (LCD) device with a stable pretilt angle has been achieved on ion-bombarded silicon nitride (SiN x) surfaces. The surface energy variation of the SiN x surface with the bonding conversion achieved via ion beam bombardment makes it possible to control the liquid crystal (LC) pretilt angle. X-ray photoelectron spectroscopy revealed that the vertical orientation of the LC molecules occurs on the SiO 2 surface and that this orientation is directionally transformed upon ion bombardment of SiN x thin films. Moreover, the upgraded electro-optical characteristics of VA-LCDs showed potential for high-performance LCD applications on modified SiN x surfaces.

This paper describes polymeric nanostructures with dynamically tunable wetting properties. Centimeter-scale areas of monolithic nanoridges can be generated by strain relief of thermoplastic polyolefin films with fluoropolymer skin layers. Changing the amount of strain results in polyolefin ridges with aspect ratios greater than four with controlled feature densities. Surface chemistry and topography are demonstrated to be able to be tailored by SF6 -plasma etching to access multiple wetting states: Wenzel, Cassie-Baxter, and Cassie-impregnating states. Reversible transitions among the wetting states can be realized in a programmable manner by cyclic stretching and reshrinking the patterned substrates without delamination and cracking.

Sum frequency generation was utilized to obtain a yellow laser with the wavelength of 578.4 nm for a probe laser of an Yb lattice clock. The output of an Nd:YAG laser with wavelength of 1319 nm and that of an Yb-fiber laser with wavelength of 1030 nm were passed through a waveguided periodically-poled lithium niobate (WG-PPLN) for sum frequency generation. It is required that the probe laser has a linewidth of the order of 1 Hz to fully resolve the Yb lattice clock transition. Thus, the linewidth of the probe laser was reduced by stabilizing the frequency to a super-cavity. This was made of ULE with a low thermal expansion coefficient, and was mounted on an active vibration-isolation table at the optimal point for the reduced sensitivity to vibration. Also, this was installed in a vacuum chamber, and the temperature was stabilized to 1 mK level. This system was installed in an acoustic enclosure to block acoustic noise. The finesse of the super-cavity was measured to be 380 000 from the photon life time of the cavity.

The aligned liquid crystals (LCs) display on SiNx thin films using ion-beam (IB) irradiation was studied with controllability of pretilt angle depending on incident energies of the IB. The LCs alignment property for the SiNx thin films were observed to verify the practical application potential. A good LCs alignment of vertical alignment LCs cells on SiNx thin film surfaces irradiated with incident IB energy of 1800eV was achieved. The atomic force microscopy (AFM) images of LCs on SiNx thin film surfaces irradiated with IB energy was used for the surface analysis.

This study was carried out to screen and identify the chitinase and chitosanase producing microorganisms from the feces of calves medicated DFM sincluding chitin in order to do the immune fortification of Korean Native calves. Ten isolates were grown in the medium containing chitin and chitosan that had more than 10 5 cfu/g in feces. Among these 10 strains, 2 strains (HANDI 110 and HANDI 309) had the chitinase activities and 2 strains (HANWOO and HANYOO) had the chitosanase activities in calves' feces. They showed no reaction in hemolysis tests by utilizing chitin and chitosan. The results from morphological, physicochemical and genetical identification indicated the HANDI 110 as a strain of Escherichia fergusonii, HANDI 309 was identified as a strain of Acinetobacter parvus, HANWOO was identified as a strain of Comamonas koreensis, and HANYOO as a strain of Chryseobacterium indologenes.

We investigated the electrical properties of polycrystalline silicon (poly-Si) thin film transistors (TFTs) employing field-enhanced solid phase crystallization (FESPC). An n+ amorphous silicon (n+ a-Si) layer was deposited to improve the contact resistance between the active Si and source/drain (S/D) metal instead of ion doping. By using C–V measurement method, we could explain the diffused phosphorous ions (P+ ions) on the channel surface caused a negatively shifted threshold voltage (VTH) of −7.81 V at a drain bias of 0.1 V, and stretched out a subthreshold swing (S) of 1.698 V/dec. This process was almost compatible with the widely used hydrogenated amorphous silicon (a-Si:H) TFT fabrication process and also offers a better uniformity when compared to the conventional laser-crystallized poly-Si TFT process because of non-laser crystallization.

Leukotriene B4 (LTB4) is a potent chemoattractant for leukocytes and considered to be an inflammatory mediator. Human BLT2 (hBLT2) is a low-affinity G-protein coupled receptor for LTB4 and mediates pertussis toxin-sensitive chemotactic cell movement. Here, we dissected the interaction between hBLT2 and G-protein alpha subunits using GST fusion proteins containing intracellular regions of hBLT2 and various Gα protein including Gα i1, Gα i2, Gα i3, Gα s1, Gα o1, and Gα z. Among the tested Gα subunits, Gα i3 showed the highest binding to the third intracellular loop region of hBLT2 with a dissociation constant (KD) of 5.0 × 10?6 M. These results suggest that Gα i3 has the highest affinity to hBLT2, and the third intracellular loop region of hBLT2 is the major component for the interaction with Gα i3.

In tissue engineering and wound-healing applications, dermal substitutes are used to provide fibroblasts with the mechanical support for their growth and then to facilitate the skin formation. In this study, three-dimensional porous poly(lactic-co-glycolic acid) (PLGA) 65/35 scaffolds were prepared and then the composites of the scaffolds and human fetal dermal fibroblasts were fabricated as a tissue-engineered dermal substitute. The function and tissue compatibility of the artificial dermal substitute were evaluated at the levels of gene expression (by RT-PCR) and protein expression (total collagen quantities), as well as by histological and immunohistochemical analysis. The PCR products indicated that the mRNA of type-I collagen, mainly secreted by the fibroblasts onto the PLGA scaffolds, was clearly expressed after 4 weeks. The amount of total collagen synthesized from the cells was shown to increase gradually during the initial culture period and slightly decreased afterwards. After 8 weeks of culture, the fibroblasts were well attached and migrated entirely throughout the pores of the PLGA scaffold with normal function. Furthermore, the positively stained type-I collagen was intensively detected throughout the pores. These results suggest that the function and tissue compatibility may be important criteria in evaluating an artificial tissue-engineered skin.

We developed an a‐Si TFT‐LCD panel with integrated gate driver circuit using a standard 5‐MASK process. To minimize the effect of the a‐Si TFT current and LC's capacitance variation with temperature, we developed a new a‐Si TFT circuit structure and minimized coupling capacitance by changing vertical architecture above gate driver circuit. Integration of gate driver circuit on glass substrate enables single chip and 3‐side free panel structure in a‐Si TFT‐LCD of QVGA (240 × 320) resolution. And using double ASG structure the dead space of TFT‐LCD panel could be further decreased.

Highly porous poly(ethylene glycol) (PEG) hydrogel scaffolds crosslinked with hydrolyzable polyrotaxane for cartilage tissue engineering were prepared by a solvent casting/salt leaching technique. The resultant scaffolds have well interconnected microporous structures ranging from 87 to 90%. Pore sizes ranging from 115.5–220.9 μm appeared to be dependent on the size of the sieved sodium chloride particulates. Moreover, a dense surface skin layer was not found on either side of the scaffold surfaces. Using microscopic Alcian blue staining of the chondrocyte-seeded scaffolds, well adhered cells and newly produced glycosaminoglycans (GAG) were confirmed. Following the initial chondrocyte seeding onto the hydrogel scaffolds, the cell number was significantly increased, reaching 149, 877, and 1228 cells/mg of tissue at 8, 15, and 21 days in culture, respectively. The micrograph shows well adhered and spread chondrocytes in the interior pores and a cartilaginous extracellular matrix with a GAG fraction produced from the chondrocytes. Results suggest that the PEG hydrogel scaffolds crosslinked with the hydrolyzable polyrotaxane are a promising candidate for chondrocyte culture. © 2003 Wiley Periodicals, Inc. J Biomed Mater Res 67A: 1087–1092, 2003

We have prepared porous polyrotaxane hydrogels by using the salt leaching technique. Porous hydrogels were found to have a uniform and highly porous structure. The size of pores in each hydrogel was directly proportional to the size of the sodium chloride particle used. Structural uniformity of the hydrogels is useful not only for uniform cell distribution, but also for well-controlled material properties. Uniform pore size and distribution may ensure the diffusion of nutrients throughout of the gel and the removal of metabolic wastes from the system. The results of an erosion study in phosphate-buffered saline showed that the erosion time of porous polyrotaxane hydrogels was controlled by the poly(ethylene glycol) (PEG) content in the hydrogels. The erosion time of the porous polyrotaxane hydrogel was observed to be almost the same with the non-porous polyrotaxane hydrogel with the same PEG content. From the erosion study, the erosion time of the polyrotaxane hydrogel may be independent of its morphology. Easy control of the erosion time in the polyrotaxane hydrogels is useful in the preparation of scaffolds for tissue engineering.

Thermodynamic analysis of inclusion complexation between α-cyclodextrin-based molecular tube (MT) and poly(ethylene oxide)-block-poly(tetrahydrofuran)-block-poly(ethylene oxide) triblock copolymer (PEO-b-PTHF-b-PEO) was carried out in terms of isothermal titration calorimetry. As for the titration of MT to PEO in aqueous solution, no strong exothermic peak was observed. On the contrary, strong exothermic peaks were observed for the titration of MT to PEO-b-PTHF-b-PEO in aqueous solution, which is attributable to inclusion complexation between MT and the triblock copolymer. Thermodynamic parameters were obtained for the process of inclusion complexation between MT and PEO-b-PTHF-b-PEO by changing the molecular weight of MT and the temperature. The stoichiometry of inclusion complexation changes in relation to the molecular weight of MT. Interestingly, it was found that MT with a large molecular weight behaves as the host molecule having multibinding sites. Thermodynamic parameters suggested that van der Wa...

Poly(propylene glycol) (PPG)-grafted dextran was synthesized by conjugating amino-terminated PPG with hydroxyl groups of dextran, and its inclusion complexation property was investigated. The average number of grafted PPG per dextran was changeable in the range of 1.1 to 38.5. The formation of inclusion complexes between the PPG grafts and β-cyclodextrins (β-CDs) and their crystalline structures were characterized by 13 C CP/MAS NMR and X-ray spectroscopies. These hydrogel systems showed a thermall reversible sol-gel transition based on supramolecular assembly and dissociation between host and guest moieties. The results of theological measurements and solgel transition temperature of the hydrogels suggested that the aggregate channel-type crystalline domain was critical to control the transition temperature in terms of initial feed molar ratio of PPG and β-CD and the graft number of PPG constituents. These thermoreversible hydrogel systems showed rapid gelation properties, which may be useful for biomedical application, especially injectable drug deliver systems.

Novel functional polymers utilizing specific host/guest interactions were designed by introducing α-CD host molecules into poly(e-lysine) chains as side groups. An interesting phase separation was observed as a result of the inclusion complexation between the polymeric host and 3-(trimethylsilyl)propionic acid as a model guest in aqueous media. This water-soluble polymeric host would be useful for various applications, particularly drug delivery, due to its biodegradability, low toxicity, and unique functionality represented as a complexation-induced phase separation.

L-Arginine was immobilized into a supramolecular-structured polyrotaxane to examine the generation of nitric oxide, with a view to improving antithrombosis and the blood compatibility of polymeric biomaterials. L-Arginine was immobilized to the hydroxyl groups of alpha-cyclodextrins in the polyrotaxane via an ester linkage, and the nitric oxide generation and L-arginine release behavior were characterized. L-Arginine-immobilized polyrotaxane was insoluble in water, but was found to generate nitric oxide when placed in Tris-HCI buffer supplemented with activators. L-Arginine-immobilized polyrotaxane exhibited sustained nitric oxide generation for a period of 250 h. L-Arginine was completely released by non-enzymatic hydrolysis from 200 h to 700 h, with a lag-time for the first 200 h. Consequently, after the generation of nitric oxide and the release of L-arginine from the L-arginine-immobilized polyrotaxane, the residual component will be a polyrotaxane with superior biocompatibility and mechanical properties. These results suggest that L-arginine-immobilized polyrotaxane can be useful in a wide range of medical applications, including use as a nitric oxide generative system for antithrombosis, coating and blending materials of hydrophobic extracorporeal circuits, and implantable catheters.

Supramolecular-structured hydrogels were prepared on basis of the inclusion complexation between poly(ethylene glycol) grafted dextrans and α-cyclodextrins (α-CDs) in aqueous media. The inclusion complexes from the PEG grafted dextrans showed a unique gel−sol phase transition which cannot be obtained from usual polymer inclusion complexes that form crystalline precipitates. The gel−sol transition was based on the supramolecular assembly and dissociation, and the transition was reversible with hysteresis. The transition temperature was controllable by variation in the polymer concentration and the PEG content in the graft copolymers as well as the stoichiometric ratio between the guest and host molecules. The properties of the hydrogel were characterized by DSC, X-ray diffraction, and 13C CP/MAS NMR. The X-ray diffraction data indicated that the gel contains a channel-type crystalline structure, demonstrated by a strong reflection at 2θ = 20° (d = 4.44 A). It was confirmed from the DSC and 13C CP/MAS NMR m...

A novel chemical modification of biological tissues was developed aimed at improving biocompatibility and calcification resistance. This method involved the additional grafting of sulfonated PEO (PEO-SO(3)) or heparin after conventional glutaraldehyde (GA) fixation of bovine pericardium (BP). The amino groups of PEO-SO(3) or heparin were utilized to react to the GA residues to block them. The PEO-SO(3) or heparin grafted tissues demonstrated a slightly higher shrinkage temperature and tensile strength, but greater resistance to collagenase digestion, than GA treated ones. These results suggest that modified tissues have improved durability due to the grafting and filling effect of PEO-SO(3) or heparin in addition to the GA cross-linking. At the direct contact cytotoxicity test in vitro, PEO-SO(3) or heparin grafted tissue was shown to be nontoxic, while relatively significant cytotoxicity was observed for the GA treated tissues, possibly due to the release of GA. From the in vivo calcification study, calcium contents deposited on the modified tissues were much less than those on GA treated tissues. Such a decreased calcification might be explained by the decrease of residual GA groups during the additional treatment, and the space-filling effect and the nonadhesive property and/or the blood compatibility of PEO-SO(3) or heparin grafted covalently. The newly modified tissue patch was observed to show improved pathological assessibility including less inflammation and tissue reactions. This simple modification method may be useful for calcification-resistant and blood-compatible tissue patches for cardiovascular implants.

Novel poly(ethylene glycol) (PEG) and sulfonated PEG (PEG-SO(3)) acrylate copolymers have been prepared and characterized to apply as coating and blending materials for biomedical applications. The modified surfaces using acrylate copolymers demonstrated increased hydrophilicity, possibly due to the hypothesized reorientation of PEG/PEG-SO(3) chains into water phase. All copolymer surfaces demonstrated less platelet adhesion than control. In addition, platelet adhesion on copolymer surfaces decreased as the chain length of PEG and sulfonated PEG in copolymers increases. All copolymer surfaces reduced bacterial adhesion significantly and the adhesion level differs depending on surfaces as well as media. The obtained results attest to the usefulness of these copolymers as a coating or additive material to improve the blood compatibility of blood contacting devices.

We fabricated a new top-gate n-type depletion-mode polycrystalline silicon (poly-Si) thin-film transistor (TFT) employing alternating magnetic-field-enhanced rapid thermal annealing. An n+ amorphous silicon (n+ a-Si) layer was deposited to improve the contact resistance between the active Si and source/drain (S/D) metal. The proposed process was almost compatible with the widely used hydrogenated amorphous silicon (a-Si:H) TFT fabrication process. This new process offers better uniformity when compared to the conventional laser-crystallized poly-Si TFT process, because it involves nonlaser crystallization. The poly-Si TFT exhibited a threshold voltage (VTH) of -7.99 V at a drain bias of 0.1 V, a field-effect mobility of 7.14 cm2/V ldr s, a subthreshold swing (S) of 0.68 V/dec, and an ON/OFF current ratio of 107. The diffused phosphorous ions (P+ ions) in the channel reduced the VTH and increased the S value.

Polyethylene oxide (PEO) surfaces were prepared by the addition of PEO- and sulfonated PEO-containing amphiphilic block copolymers as surface-modifying additives in a segmented polyurethane (PU). PEO-PPO-PEO triblock copolymers (Pluronics) with different PEO chain lengths (from 2 to 80) were used as additives. The prepared film surfaces were characterized by the measurement of dynamic water contact angles and electron spectroscopy for chemical analysis. It was observed that the PU films containing 10 wt % of PEO additives were surface-saturated with the additives regardless of their PEO chain length, but the PEO chains were more projected from the film surfaces containing the additives with longer PEO chains. The water absorption of the films increased largely with the increasing PEO chain length of the additives. The addition of PEO additives produced film surfaces that were in a gel-like state. The films demonstrated some extraction of the PEO additives. However, the additives with higher molecular weights were entrapped more stably into the PU matrix. The mechanical properties (tensile strength and elongation) of the films were changed by the addition of PEO additives, but the differences were not significant compared to the control PU. The platelet adhesion on the film surfaces decreased with increasing PEO chain length of the additives. The film surface containing additives with long PEO chains (chain length of 80) was particularly effective in preventing platelet adhesion. The effect of negatively charged sulfonate groups on the prevention of platelet adhesion appeared only on the film surfaces containing additives with short PEO chains. For longer PEO chains, the chain mobility effect was more dominant than the negative charge effect on the prevention of platelet adhesion. © 1998 John Wiley & Sons, Inc. J Biomed Mater Res, 40, 314–323, 1998.

Uridinediphospho-N-acetylglucosamine enolpyruvyl transferase (MurA, E.C. 2.5.1.7) is an essential bacterial enzyme that catalyzes the first step of the cell wall biosynthetic pathway, which involves the transfer of an enolpyruvyl group from phosphoenolpyruvate to uridinediphospho-Nacetylglucosamine. In this study, novel inhibitors of Haemophilus influenzae MurA (Hi MurA) were identified using high-throughput screening of a chemical library from the Korea Chemical Bank. The identified compounds contain a quinoline moiety and have much lower effective inhibitory concentrations (IC(50)) than fosfomycin, a wellknown inhibitor of MurA. These inhibitors appear to covalently modify the sulfhydryl group of the active site cysteine (C117), since the C117D mutant Hi MurA was not inhibited by these compounds and excess dithiothreitol abolished their inhibitory activities. The increased mass value of Hi MurA after treatment with the identified inhibitor further confirmed that the active-site cysteine residue of Hi MurA is covalently modified by the inhibitor.

Non-structural protein 1 (NS1) of the influenza A virus (IAV) inhibits the host's innate immune response by suppressing the induction of interferons (IFNs). Therefore, blocking NS1 activity can be a potential strategy in the development of antiviral agents against IAV infection. In the present study, we selected a single-stranded DNA aptamer specific to the IAV NS1 protein after 15 cycles of systematic evolution of ligands by exponential enrichment (SELEX) procedure and examined the ability of the selected aptamer to inhibit the function of NS1. The selected aptamer binds to NS1 with a Kd of 18.91±3.95nM and RNA binding domain of NS1 is determined to be critical for the aptamer binding. The aptamer has a G-rich sequence in the random sequence region and forms a G-quadruplex structure. The localization of the aptamer bound to NS1 in cells was determined by confocal images, and flow cytometry analysis further demonstrated that the selected aptamer binds specifically to NS1. In addition, luciferase reporter gene assay, quantitative RT-PCR, and enzyme-linked immunosorbent assay (ELISA) experiments demonstrated that the selected aptamer had the ability to induce IFN-β by suppressing the function of NS1. Importantly, we also found that the selected aptamer was able to inhibit the viral replication without affecting cell viability. These results indicate that the selected ssDNA aptamer has strong potential to be further developed as a therapeutic agent against IAV.

We introduce the characteristics of the titanium dioxide (TiO(2)) inorganic film deposited by rf magnetron sputtering for liquid crystal display applications. The TiO(2) films demonstrated vertical alignment (VA) of the liquid crystals (LCs) obtained by using ion-beam (IB) bombardment. As observed by using x-ray photoelectron spectroscopy, the chemical structure of the TiO(2) was changed by IB bombardment, altering the Ti-O bonding of the Ti 2p spectra to lower intensity levels. Breaking Ti-O bonding by IB bombardment created pretilt angles between the TiO(2) film and LC molecules. The better voltage-transmittance characteristics of the VA LCDs based on TiO(2) film were measured and compared with the same characteristics of polyimide film.

Bacterial UDP-N-acetylglucosamine enolpyruvyl transferase (MurA) catalyzes the transfer of enolpyruvate from phosphoenolphyruvate (PEP) to uridine diphospho-N-acetylglucosamine (UNAG), which is the first step of bacterial cell wall synthesis. We identified thimerosal, thiram, and ebselen as effective inhibitors of Heamophilus influenzae MurA by screening a chemical library that consisted of a wide range of bioactive compounds. When MurA was preincubated with these inhibitors, their 50% inhibitory concentrations (IC50s) were found to range from 0.1 to 0.7 microM. In particular, thimerosal suppressed the growth of several different Gram-negative bacteria such as Escherichia coli, Pseudomonas aeruginosa, Salmonella typhimurium at a concentration range of 1-2 microg/ml. These inhibitors covalently modified the cysteine residue near the active site of MurA. This modification changed the open conformation of MurA to a more closed configuration, which may have prevented the necessary conformational change from occurring during the enzyme reaction.

Electro-optic performance of a liquid-crystal (LC) system is enhanced by TiO(2) nanoparticle dispersed in nematic liquid crystal (NLC). The 2.5 V threshold voltage of LC for device operation is lowered to 0.5 V through TiO(2) nanoparticle mixing up to 2 wt.%. To characterize the shape and size distribution of the nanoparticles, high-resolution transmission electron microscopy is employed. Transmittance spectra for the TiO(2) dispersed LC structure and nondispersed LC structure showed that transparency of the TiO(2) dispersed LC is similar to that of pure liquid LC.

We have fabricated the new top gate depletion mode n-type alternating magnetic field enhanced rapid thermal annealing (AMFERTA) polycrystalline silicon (poly-Si) thin film transistors (TFTs), which show the excellent electrical characteristics and superior stability compared with hydrogenated amorphous silicon (a-Si:H) TFTs and excimer laser crystallized (ELC) low temperature polycrystalline silicon (LTPS) TFTs. The fabricated AMFERTA poly-Si TFTs were not degraded under hot-carrier stress, and highly biased vertical field stress. The considerably large threshold voltage shift (ΔVTH) and trap state density reducing were occurred when the gate bias and drain bias were both large enough. The dominant mechanism of instability in the fabricated depletion mode AMFERTA poly-Si TFTs may be due to carrier induced donor-like defects reduction within the channel layer, especially near the drain junction.

Sulfonated polyrotaxanes (PRx-SO(3)'s), in which sulfonated alpha-cyclodextrins (alpha-CDs) were threaded onto the poly(ethylene glycol) (PEG) segments in a PEG-b-poly(propylene glycol) (PPG)-b-PEG triblock copolymer (Pluronic) capped with benzyloxycarbonyl (Z)-L-phenylalanine (Z-L-Phe), were prepared as a novel surface-modifying biomaterial. Surface modification of the polyurethane (PU) was carried out by blending the PRx-SO(3)'s with a PU solution, followed by solution casting. The incorporated PRx-SO(3)'s led to the enhanced hydrophilicity by changing the surface properties of the PU matrix. Modified PUs showed the stable entrapment of the PRx-SO(3)'s with little extraction into water and enhanced mechanical properties after exposure to water compared to the PU control. The incorporated PRx-SO(3)'s repelled the proteins and kept them from closely approaching the surface areas, prevented platelet activation by thrombin, and effectively repelled bacteria. These results suggest that both the supramolecular structure of the polyrotaxanes and exposure of the sulfonated groups onto the surfaces contribute to these phenomena. Thus, surface modification with PRx-SO(3)'s is suggested to be useful for the fabrication of biocompatible medical devices.

Polyrotaxanes, in which α-cyclodextrins (α-CDs) are threaded onto poly(ethylene glycol)-b-poly(propylene glycol)-b-poly(ethylene glycol) triblock copolymers (Pluronic) capped with benzyloxycarbonyl(Z)-L-phenylalanine (Z-L-Phe), were prepared, and sulfopropyl groups were introduced to hydroxyl groups of α-CDs in the polyrotaxanes. The supramolecular structure and the chemical composition of the polyrotaxanes after the sulfonation were confirmed by 1H-NMR, 13C-NMR, and elemental analysis. Anticoagulant activity of the polyrotaxanes and sulfonated polyrotaxanes was measured by activated partial thromboplastin time (APTT). It was found that the polyrotaxanes and the sulfonated polyrotaxanes showed greater anticoagulant activity than Pluronic itself, suggesting that both the supramolecular structure of the polyrotaxanes and the sulfonated groups contribute to the inhibition of intrinsic coagulation factors. Finally, our designed polyrotaxanes are suggested to be a promising candidate when fabricating blood-compatible medical devices by blending with or coating on clinically used polymers. © 2002 Wiley Periodicals, Inc. J Biomed Mater Res 60: 186–190, 2002

Biological porcine tissue was modified by the direct coupling of sulphonated poly(ethylene oxide) (PEO-SO3) containing amino acid end groups after glutaraldehyde fixation. The calcification of the modified tissue [bioprosthetic tissue (BT)-PEO-SO3] and control (BT control) was investigated by in vivo rate subdermal, canine aorta-illiac shunt and right ventricle-pulmonary artery shunt implantation models. Less calcium deposition of BT-PEO-SP3 than of BT control was observed in in vivo tests. Such a reduced calcification of BT-PEO-SO3 can be explained by decreases of residual glutaraldehyde groups, a space filling effect and, therefore, improved biostability and synergistic blood-compatible effects of PEO and SO3 groups after the covalent binding of PEO-SO3 to tissue. This simple method can be a useful anti-calcification treatment for implantable tissue valves.

Polyurethane (PU) surfaces were modified by coupling polyethylene glycol (PEG; molecular weight, 1,000) chains carrying different terminal groups (PU-PEG1K-OH, PU-PEG1K-NH2, PU-PEG1K-SO3) and longer PEG chains (MW, 3,350; PU-PEG3.4K-OH). The modified PU surfaces have the same PEG (1K) chain density. Surface induced platelet activation was evaluated by measuring cytoplasmic free calcium concentration in platelets contacting modified surfaces, and platelet adhesion onto modified surfaces was investigated in vitro. Cytoplasmic free calcium levels in platelets contacting PU-PEG-SO3 remained relatively constant, in contrast to the significant increase observed for PU-PEG-NH2, PU-PEG-OH, and control PU surfaces. The degree of platelet adhesion clearly demonstrates that all PEG graft surfaces prevented platelet adhesion. Among PEG1K surfaces, PU-PEG-SO3 shows the lowest platelet adhesion. In the case of relatively longer PEG grafted surfaces (PU-PEG3.4K-OH and PU-PEG3.4K-Hep), both surfaces were found to prevent the increase in both cytoplasmic free calcium and platelet adhesion. These results suggest that longer PEG chain grafting is more effective than shorter grafting in preventing platelet activation and adhesion because of the highly dynamic movement of hydrated PEG chains at the interface. In addition, in vitro platelet interaction is dependent upon terminal groups of PEG chains on PEG1K series surfaces.

Rigid polymers have been of great interest in recent years because of their outstanding mechanical properties and potential applications for the high performance materials. The rigid polymers, however, are almost nonprocessible and structural modifications to improve processibility have been the subject of investigation thereafter. Attempts were made by introducing the flexible linkages or nonlinear aromatic moieties in the polymer main chain1–7 and by coupling of flexible side chains.8–13 Attachment of flexible alkoxy side chains to the several rigid polymers were proven to be successful to lower the transition temperatures and to enhance the solubility of polymers. The flexible side chains apparently act as a bound solvent, also yielding a layered mesophase of the polymer chains. The layered mesophase and systematic changes with side chain length also have been of interest in these rigid polymers. In this work we prepared polyamides having bulky rigid moiety in the main chain and alkoxy side chains. We report the synthesis and properties as well as molecular packing behavior associated with the side chains.

We have fabricated p-channel metal oxide semiconductor (PMOS) solid-phase crystallization silicon (SPC-Si) thin-film transistors (TFTs) employing an alternating magnetic field on a glass substrate, which exhibit better current uniformity than excimer-laser annealed low-temperature polycrystalline silicon (ELA LTPS) TFTs, and superior stability to hydrogenated amorphous silicon (a-Si:H) TFTs. The leakage current of the SPC-Si TFT was larger than that of an a-Si:H TFT and an ELA poly-Si TFT owing to its many grain boundaries. The leakage current of the SPC-Si TFT was successfully suppressed through off-state-bias annealing. Off-state-bias annealing forced electron carriers to be trapped in the gate insulator, thus reducing the gate–drain field and suppressing the leakage current of the SPC-Si TFT. Our experimental result showed that the trapped electron charges in the gate insulator did not escape under thermal annealing or AC-bias stress. The off-state-bias annealed PMOS SPC-Si TFT could be a suitable pixel element for high-quality display applications employing an alternating magnetic field.

Introduction The polycrystalline silicon (poly-Si) thin-film transistors (TFTs) employing alternating magnetic-fieldenhanced thermal annealing (AMFERTA) have been increasingly attracted for pixel elements of active matrix organic light emitting-diode (AMOLED) displays due to good uniformity and large-area applicability [1],[2]. Recently, the bottom-gate poly-Si TFTs have much interested because the fabrication process are almost identical to those of widely used amorphous silicon (a-Si) TFTs for manufacturing of TFT-LCD [3]. Therefore, the bottom-gate TFTs can be applicable for the low-cost and large-size flat panel displays. The purpose of this paper is to report for the first time about the characteristics of hot-carrier stressed bottomgate AMFERTA poly-Si TFTs. It can be contributed to understand the bottom-gate AMFERTA TFTs and to enlarge low temperature poly-Si TFT applications.