A Comprehensive Study of the Interaction between Peptidoglycan Fragments and the Extracellular Domain of Mycobacterium tuberculosis Ser/Thr Kinase PknB

The Mycobacterium tuberculosis Ser/Thr kinase PknB is implicated in the regulation of bacterial cell growth and cell division. The intracellular kinase function of PknB is thought to be triggered by peptidoglycan (PGN) fragments that are recognized by the extracytoplasmic domain of PknB. The PGN in the cell wall of M. tuberculosis has several unusual modifications, including the presence of N-glycolyl groups (in addition to N-acetyl groups) in the muramic acid residues and amidation of d -Glu in the peptide chains. Using synthetic PGN fragments incorporating these diverse PGN structures, we analyzed their binding characters through biolayer interferometry (BLI), NMR spectroscopy, and native mass spectrometry (nMS) techniques. The results of BLI showed that muropeptides containing 1,6-anhydro-MurNAc and longer glycan chains exhibited higher binding potency and that the fourth amino acid of the peptide stem, d -Ala, was crucial for protein recognition. Saturation transfer difference (STD) NMR spectroscopy indicated the major involvement of the stem peptide region in the PASTA-PGN fragment binding. nMS suggested that the binding stoichiometry was 1:1. The data provide the first molecular basis for the specific interaction of PGN with PknB and firmly establish PGNs as the effective ligands of PknB.     

Preparation of silk fibroin and polyallylamine composites

Composite films made of silk fibroin (SF) and polyallylamine (PAA) are prepared that contain various compositional ratios. These materials are analyzed to elucidate the resulting physical properties and to assess their potential toward advanced applications as industrial materials. The composite films are obtained from a SF and PAA binary system by dry casting from aqueous solution. These composite films exhibit excellent processability such as film forming capabilities, and the elongation at break is increased in the wet state. The differential scanning calorimetry (DSC) curves of the composites suggest that a mutual interaction takes place between the SF and PAA. This interaction is believed to occur because the endothermic peak, corresponding to the individual polymer, shifts with increasing SF content. The random coil conformation of the SF is present, regardless of the PAA blending, as confirmed by FTIR and DSC measurements. Additionally, living cells from Antheraea pernyi and Bombyx mori insect tissues are shown to grow effectively on the composite films. Maximum growth levels occur when the cultivation flask is coated with the material in SF/PAA ratios of 75:25 to 25:75. © 2002 Wiley Periodicals, Inc. J Appl Polym Sci 84: 1963–1970, 2002; DOI 10.1002/app.10491     

Heavy oil sorption and recovery by using carbon fiber felts

On fibrous carbon materials, including activated carbon fibers, sorption capacity for heavy oils, less viscous A-grade and more viscous C-grade, was determined. Sorption capacity depended strongly on their bulk density; the correlation was the same as that found previously on exfoliated graphite and carbonized fir fibers. On carbon fiber felts, excellent recycling performance was observed, though sorption capacity was not so high as on exfoliated graphite and carbonized fir fibers. By filtration under suction, about 90% of sorbed A-grade heavy oil could be recovered and no decrease in sorption capacity was detected even after eight cycles. By washing with solvents, n-hexane for A- and C-grade oils and A-grade oil for C-grade oil, almost 100% recovery with no marked reduction in sorption capacity was found for each cycle. For the felts of PAN-based carbon fibers, rather severe operations for oil recovery, centrifugation and squeezing with twisting, could be applied without pronounced decreases in sorption capacity and recovery ratio.     

An absolute requirement of fructose 1,6-bisphosphate for the Lactobacillus caseiL-lactate dehydrogenase activity induced by a single amino acid substitution

Lactobacillus casei allosteric L-lactate dehydrogenase (L-LDH)absolutely requires fructose 1,6-bisphosphate [Fru(1,6)P2] forits catalytic activity under neutral conditions, but exhibitsmarked catalytic activity in the absence of Fru(1,6)P₂ underacidic conditions through the homotropic activation effect ofsubstrate pyruvate. In this enzyme, a single amino acid replacement,i.e. that of His205 conserved in the Fru(1,6)P₂-binding siteof certain allosteric L-LDHs of lactic acid bacteria with Thr,did not induce a marked loss of the activation effect of Fru(1,6)P₂or divalent metal ions, which are potent activators that improvethe activation function of Fru(1,6)P₂ under neutral conditions.However, this replacement induced a great loss of the Fru(1,6)P₂-independentactivation effect of pyruvate or pyruvate analogs under acidicconditions, consequently indicating an absolute Fru(1,6)P₂ requirementfor the enzyme activity. The replacement also induced a significantreduction in the pH-dependent sensitivity of the enzyme to Fru(1,6)P₂,through a slight decrease and increase of the Fru(1,6)P₂ sensitivityunder acidic and neutral conditions, respectively, indicatingthat His205 is also largely involved in the pH-dependent sensitivityof L.casei L-LDH to Fru(1,6)P₂. The role of His205 in the allostericregulation of the enzyme is discussed on the basis of the knowncrystal structures of L-LDHs.     

Rapid and Continuous Hydrothermal Synthesis of Boehmite Particles in Subcritical and Supercritical Water

Hydrothermal synthesis of AlOOH particles from an Al(NO₃)₃ aqueous solution was examined in subcritical and supercritical water. Continuous and rapid production of AlOOH fine particles was achieved by mixing a metal salt aqueous solution with preheated water fed from another line. The effects of temperature, pressure, and initial concentration of solution on the particle size, morphology, and crystal structure of the metal (hydrous) oxides were examined. These properties are strongly affected by slight changes in pressure and temperature. This strong effect is attributed to the large changes in the properties of water near its critical point resulting from the slight changes in pressure and temperature. The results suggest the simultaneous control of size, crystal structure, and morphology of the particles by hydrothermal synthesis in supercritical water.     

ARCOR,a new photolithography technique with antireflective coating on resist

The multiple interference effect is one of the major causes of the fluctuation in critical dimension control (CD) and in mark detection for alignment. Suppressing this effect is critical for future photolithography. We propose a new photolithography technique called anti reflective coating on resist (ARCOR), which improves linewidth accuracy and overlay accuracy by suppressing multiple interference. ARCOR consists of relatively simple processes: A clear antireflective film is spun onto the resist prior to the mark detecting for alignment and exposure. The film is subsequently removed and the resist developed in the conventional way. ARCOR differs from ARC, which suppresses the reflection at the resist/substrate interface. ARCOR suppresses the reflection at the air/resist interface. ARCOR allows mark detection and exposure without light intensity-loss and multiple interference. The experiments mainly examine polysiloxane and perfluoroalkylpolyether as ARCOR materials. It is shown that linewidth accuracy can be improved from 0.3 to 0.03 μm. The signal-to-noise ratio of the alignment signal is drastically improved, and the overlay error is about half that of the conventional method. ARCOR is also effective for directly measuring the reflectivity at the resist/substrate interface, which is a key parameter of the multiple interference effect and the halation. Using ARCOR and a thin resist film, the measured ratio of reflected light to incident light indicates the reflectivity at the resist/substrate interface. Because, the probe light does not reflect off the resist surface and the intensity-loss at the resist surface is suppressed. With perfluoroalkylpolyether film, the measurement error is ～ 1.5%.     

Preparation of water‐repellent silks by a reaction with octadecenylsuccinic anhydride

Silk fibers and membranes were acylated with octadecenylsuccinic anhydride (ODSA) at 75°C for different times. Swelling [N,N‐dimethylformamide (DMF) and dimethyl sulfoxide (DMSO)] and nonswelling (xylene) solvent media were used for the reaction. Silk membranes that reacted in DMF or DMSO displayed faster reaction kinetics and attained higher weight‐gain values than fibers. The effect of the solvent on the reaction yield was in the following order: DMSO > DMF ? xylene. The Fourier transform infrared spectra of acylated silk samples showed the characteristic absorption bands of the anhydride at 2990, 2852, 1780–1700, and 1170 cm^?1. The intensity of the latter band, which increased linearly with the weight gain, was used as a marker for evaluating the reaction kinetics of the samples acylated in the nonswelling medium. The moisture regain and water retention of silk fibers acylated with ODSA decreased significantly, regardless of the solvent system used. Accordingly, the water repellency increased. Acylation induced an increase in the thermal stability of the silk fibers and membranes. Fine particles adhering to the surfaces of the silk fibers acylated in xylene were detected by scanning electron microscopy. © 2003 Wiley Periodicals, Inc. J Appl Polym Sci 89: 324–332, 2003     

Role of network nematicity in swelling and phase equilibria of polymer networks in nematic solvents

Swelling and phase equilibria of polymer networks in a low molecular mass liquid crystal (LC) have been investigated as a function of network nematicity. LC networks with varying nematicity were prepared by copolymerization of mixtures of mesogenic monomers and non-mesomorphic (styrene) monomers with various compositions. Molar fraction of mesogenic monomer (x) in copolymer network strongly influences the swelling behavior as well as the nematic-isotropic (N-I) transitions in both dry and swollen states. The swollen networks of sufficiently high x with strong nematicity exhibit a sharp N-I transition and simultaneously undergo a discontinuous change in gel volume, i.e. volume phase transition. Meanwhile, the swollen networks of x≤0.8 with less nematicity show a broad N-I transition, and the resulting volume change proceeds continuously over a finite temperature range. When x decreases further down to less than 0.5, the nematicity of the dry networks vanishes. The nematic ordering in the swollen copolymer networks of x<0.5 occurs at the same temperature as the N-I transition temperature of the pure nematic solvent (T_NI^S), which yields the inflection in the swelling-temperature curve. A mean field theory considering network nematicity as a variable describes the effects of x on volume phase transition such as a shift of T_NI^G and a change in the magnitude of volume transition, apart from the emergence of continuous volume transition due to the broad N-I transition. The purely isotropic network of x=0 appreciably swells in the nematic solvent. The solvent inside the gel forms the nematic phase at the temperatures below which conflicts with the classical theoretical prediction that nematic ordering of LC solvent in fully swollen isotropic network never occurs.     

Hydrogenation of nitro compounds with supported platinum catalyst in supercritical carbon dioxide

Hydrogenation of a series of substituted nitro compounds such as 2-,3-,4-nitroanisole, 2-,3-,4-nitrotoluene, 2,4-dinitrobenzene and 2,4-dinitrotoluene has been studied in supercritical carbon dioxide, scCO₂ (two phases), and ethanol (three phases) with a 5 wt.% carbon supported platinum catalyst. The solubility of these compounds in scCO₂ has also been examined in the presence and absence of hydrogen. The solubility of those nitro compounds increases with increasing CO₂ pressure but decreases with the presence of hydrogen. The solubility is in the order of nitrotoluene > nitroanisole > dinitrotoluene, dinitrobenzene. Although the total conversion obtained with hydrogenation in scCO₂ is similar to that in ethanol, the selectivity to amino products is higher in the former reaction medium, indicating that scCO₂ is an ideal medium for the production of amino compounds with hydrogenation of nitro substrates using conventional supported metal catalysts.     

Absorption of metal cations by modified B. mori silk and preparation of fabrics with antimicrobial activity

Silk fabrics were modified by treatment with tannic acid (TA) solution or by acylation with ethylenediaminetetraacetic (EDTA) dianhydride. Kinetics of modification with TA and acylation with EDTA–dianhydride was investigated. The physico‐mechanical properties of silk fabrics acylated with EDTA–dianhydride remained unchanged regardless of chemical modification. The absorption of metal cations (Ag⁺, Cu²⁺) by untreated and modified silk fabrics was studied as a function of the kind of modifying agent, weight gain, and pH of the metal solution. The absorption of Cu²⁺ at alkaline pH was not significantly influenced by chemical modification of the silk substrate. The absorption of Ag⁺ by acylated silk remained at a level as low as untreated silk, while was enhanced by TA. The higher the content of TA, the higher the absorption of Ag⁺. With respect to the pH of the metal solution, the acylation with EDTA–dianhydride enabled silk to absorb and bind metal cations even in the acidic and neutral pH range, where tannic acid had no effect. Medium to high levels of metal desorption were exhibited by untreated and modified silk fabrics towards the metal cations, with the only exception of the silk–tannic acid–Ag complex, which displayed an extraordinary stability. All metal‐containing silks exhibited significant antibacterial activity. © 2001 John Wiley & Sons, Inc. J Appl Polym Sci 80: 297–303, 2001