Colored Fluorescent Silk Made by Transgenic Silkworms

Silk is a protein fiber used to weave fabrics and as a biomaterial in medical applications. Recently, genetically modified silks have been produced from transgenic silkworms. In the present study, transgenic silkworms for the mass production of three colors of fluorescent silks, (green, red, and orange) are generated using a vector originating from the fibroin H chain gene and a classical breeding method. The suitability of the recombinant silks for making fabrics is investigated by harvesting large amounts of the cocoons, obtained from rearing over 20 thousand silkworms. The application of low temperature and a weakly alkaline solution for cooking and reeling enables the production of silk fiber without loss of color. The maximum strain tolerated and Young's modulus of the fluorescent silks are similar to those of ordinary silk, although the maximum stress value of the recombinant silk is slightly lower than that of the control. Fabrics with fluorescent color are demonstrated using the recombinant silk, with the color persisting for over two years. The results indicate that large amounts of genetically modified silk can be made by transgenic silkworms, and the silk is applicable as functional silk fiber for making fabrics and for use in medical applications.     

Dark-field transmission electron microscopy for a tilt series of ordering alloys: toward electron tomography

Here we show a technique to obtain a tilt series of dark-field (DF) transmission electron microscopy (TEM) images in ordering alloys for tomographic three-dimensional (3D) observations. A tilt series of DF TEM images of D1a-ordered Ni4Mo precipitates in a Ni-Mo alloy was successfully obtained by adjusting a diffraction condition for a superlattice reflection from the Ni4Mo precipitates. Since the superlattice reflection usually has a long extinction distance, dynamic diffraction effects such as thickness fringes can be suppressed to some extent with precise realignment of the diffraction condition. By using the tilt series of the DF TEM images, we attempted a computed TEM tomography to visualize 3D shapes and positions of the precipitates.     

Low-Temperature Physical Properties of Ba8Ni x Ge46?x (x?=?3,?4,?6)

Polycrystalline samples of type?I clathrate Ba₈Ni _x Ge_46?x (x?=?3, 4, 6) have been synthesized using a radio-frequency (RF) induction furnace. Ba₈Ni _x Ge_46?x samples show metallic-like behavior (d??/dT?>?0) with high resistivity at room temperature, and diamagnetic susceptibility at 2?K and 300?K. The charge carriers vary from n type to p type depending on the Ni stoichiometry. The carrier concentrations at 300?K are calculated to be 5.84(3)?e^?/cell for x?=?3, 2.29(1)?e^?/cell for x?=?4, and 3.29(1)?h⁺/cell for x?=?6. The deviation of the carrier concentrations from the values expected based on 4-bonded Ni suggests that vacancies may play a very important role in the electronic states. The effective carrier masses are estimated from the low-temperature heat capacity data.     

Chemical and Physical Sensing by Organic Field‐Effect Transistors and Related Devices

Organic semiconductor films are susceptible to noncovalent interactions, trapping and doping, photoexcitation, and dimensional deformation. While these effects can be detrimental to the performance of conventional circuits, they can be harnessed, especially in field‐effect architectures, to detect chemical and physical stimuli. This Review summarizes recent advances in the use of organic electronic materials for the detection of environmental chemicals, pressure, and light. The material features that are responsible for the transduction of the input signals to electronic information are discussed in detail.     

Flexible organic transistors and circuits with extreme bending stability

Flexible electronic circuits are an essential prerequisite for the development of rollable displays, conformable sensors, biodegradable electronics and other applications with unconventional form factors. The smallest radius into which a circuit can be bent is typically several millimetres, limited by strain-induced damage to the active circuit elements. Bending-induced damage can be avoided by placing the circuit elements on rigid islands connected by stretchable wires, but the presence of rigid areas within the substrate plane limits the bending radius. Here we demonstrate organic transistors and complementary circuits that continue to operate without degradation while being folded into a radius of 100 μm. This enormous flexibility and bending stability is enabled by a very thin plastic substrate (12.5 μm), an atomically smooth planarization coating and a hybrid encapsulation stack that places the transistors in the neutral strain position. We demonstrate a potential application as a catheter with a sheet of transistors and sensors wrapped around it that enables the spatially resolved measurement of physical or chemical properties inside long, narrow tubes.     

Potential of predominant activated sludge bacteria as recipients in conjugative plasmid transfer

We investigated the possibility of conjugative plasmid transfer to the predominant bacteria in activated sludge and the factors influencing the transfer frequency in the activated sludge process. We performed conjugative transfers of a self-transmissible, broad-host-range plasmid RP4 from Escherichia coli C600 to activated sludge bacteria by broth mating. Most of the activated sludge bacteria tested could acquire plasmid RP4, although the transfer frequencies varied from 8.8 x 10(-7) to 1.3 x 10(-2) transconjugants per recipient. The transfer frequencies in several strains were similar to, or higher than, that in intraspecific transfer to E. coli HB101. Matings under various environmental conditions showed that factors relevant to physiological activity, such as temperature and nutrient conditions, seemed to affect the transfer frequency. In addition, conjugative transfer was detected even in filtered raw and treated wastewaters. Thus, the predominant activated sludge bacteria seem to have sufficient potential as recipients in conjugative plasmid transfer under the conditions likely to occur in the activated sludge process. Transfer frequency was reduced by agitation in the presence of suspended solid. This may suggest that conjugative plasmid transfer is physically inhibited in aeration tanks.     

Antimicrobial susceptibilities of Salmonella isolates obtained from layer chicken houses on a commercial egg-producing farm in Japan, 1997 to 2002

Susceptibility to antimicrobial agents was examined for 325 isolates of Salmonella enterica serotypes Cerro, Infantis, Livingstone, and Montevideo isolated from layer houses on a commercial egg-production farm in the western region of Japan between 1997 and 2002. No antimicrobials were used for therapeutic purposes on the farm during this period. From 1.8 to 3.1% of the isolates were resistant to ampicillin, chloramphenicol, and tetracycline. Resistance to streptomycin and sulfisoxazole was found in 52.9 and 65.5%, respectively, of Salmonella Montevideo isolates and in 0 to 13.2% of the isolates of the other serotypes. All the streptomycin-resistant isolates of Salmonella Montevideo also exhibited resistance to sulfisoxazole. Salmonella Montevideo isolates were first isolated in 1998, and 80.0% of the isolates obtained in this year were resistant to streptomycin and sulfisoxazole. The results suggest that Salmonella Montevideo isolates that had already acquired resistance may have been introduced into the layer houses, although the route and vehicle of transmission were uncertain. The proportion of Salmonella Montevideo isolates resistant to streptomycin and sulfisoxazole significantly decreased (P < 0.01) from 79.5% for 1998 through 1999 to 37.3% for 2000 through 2002. This decrease probably was due to the fact that no antimicrobials were used on the farm. Among 10 isolates from different serotypes obtained from 1997 to 2002 that were resistant to ampicillin, two and five isolates harbored 42- and 63-kb R plasmids and identical DraI restriction enzyme digest patterns, respectively, and carried the blaTEM gene. The results suggest that the ampicillin resistance determinants were transferred among different serotypes of Salmonella in the layer houses.     

Strain selection and scale-up fermentation for FR901379 acylase production by Streptomyces sp. no. 6907

Micafungin (FK463) is a widely used treatment for life-threatening, deep-seated fungal infections. It is an echinocandin-like lipopeptide derived from the chemical modification of deacylated FR901379, a type of lipopeptide antibiotic produced by Coleophoma empetri F-11899. The palmitoyl moiety of FR901379 is deacylated by FR901379 acylase produced by Streptomyces sp. no. 6907. In this study, our goal was to generate an improved strain of Streptomyces sp. no. 6907 capable of hyperproducing the FR901379-acylase enzyme. To accomplish this goal, modified strains of Streptomyces sp. no. 6907 were generated using UV-irradiation mutagenesis, and strain selection was performed using an agar-plate screening method to efficiently select an acylase-hyperproducing strain. Three marker indices were shown to correlate with elevated acylase production: decreased candidacidal activity of FR901379, decreased proteolytic activity on skim milk, and phenotypic characteristics. Cloning and subsequent sequencing of the acylase gene from the hyperproducing mutant revealed no mutations in either the acylase structural gene or the 5'-flanking region required for gene expression. The growth medium was also modified to maximize acylase production. We successfully increased acylase activity approximately 65-fold, compared with the original growth conditions (wild strain cultured in the original unmodified medium). To minimize formation of excess foam during the fermentation process, we optimized the parameters of agitation speed, as calculated from the discharge flow rate. Using our improved strain and the optimized medium and growth conditions, we have developed an improved and highly reproducible method for stable large-scale production of FR901379-acylase.     

Application of femtosecond laser ablation for detaching grown protein crystals from glass capillary tube

We developed a novel technique for detaching protein crystals from glass capillary tube using the counter diffusion crystallization technique by femtosecond laser irradiation. X-ray diffraction analysis demonstrated that femtosecond laser irradiation has little effect on crystallinity. This technique will contribute to progress in structural genomics as a powerful tool.     

Sweeteners interacting with the transmembrane domain of the human sweet-taste receptor induce sweet-taste synergisms in binary mixtures

Sweet enhancing effect of neohesperidin dihydrochalcone (NHDC) or cyclamate has been reported to be synergistic in human sensory tests. However, little is known about whether these synergisms are caused by the mechanism mediated by the human sweet-taste receptor. Here, we examined the sweetness intensity of sweet tastant mixtures by measuring the responses of cultured cells stably expressing the human sweet-taste receptor. The results showed that the cell response to sucrose was synergistically potentiated by the addition of NHDC or cyclamate. Moreover, a point mutation in the transmembrane domain of hT1R3 almost completely eliminated the enhancing effects of NHDC and cyclamate. These results suggest that ligand–receptor interactions in the transmembrane domain of hT1R3 are necessary for NHDC and cyclamate to elicit the synergistic potentiation of the receptor activation. Our results may provide the foundation of a molecular basis for receptor-based synergisms of sweet tastes in mixtures of diverse sweet substances.