Application of inorganic phosphate limitation to efficient isoprene production in Pantoea ananatis.

AIMS: Establishment of an efficient isoprene fermentation process by adopting inorganic phosphate limitation as the trigger to direct metabolic flux to the isoprene synthetic pathway. METHODS AND RESULTS: We constructed isoprene-producing strains of Pantoea ananatis (a member of the Enterobacteriaceae family) by integrating a heterologous mevalonate pathway and a metabolic switch that senses external inorganic phosphate (Pi) levels. This metabolic switch enabled dual-phase isoprene production, where the initial cell growth phase under Pi-saturating conditions was uncoupled from the subsequent isoprene production phase under Pi-limiting conditions. In fed-batch fermentation using our best strain (SWITCH-PphoC/pIspSM) in a 1-l bioreactor, isoprene concentration in the off-gas was maintained between 300 and 460 ppm during the production phase and at 20 ppm during the cell growth phase, respectively. The strain SWITCH-PphoC/pIspSM produced totally 2·5 g l-1 of isoprene from glucose with a 1·8% volumetric yield in 48 h. CONCLUSIONS: This proof-of-concept study demonstrated that our Pi-dependent dual-phase production system using a P. ananatis strain as a producer has potential for industrial-scale isoprene fermentation. SIGNIFICANCE AND IMPACT OF THE STUDY: This Pi-dependent dual-phase fermentation process could be an attractive and economically viable option for the production of various commercially valuable isoprenoids.

Defect structures in nematic liquid crystals around charged particles.

We numerically study the orientation deformations in nematic liquid crystals around charged particles. We set up a Ginzburg-Landau theory with inhomogeneous electric field. If the dielectric anisotropy epsilon 1 is positive, Saturn-ring defects are formed around the particles. For epsilon 1< 0 , novel "ansa" defects appear, which are disclination lines with their ends on the particle surface. We find unique defect structures around two charged particles. To lower the free energy, oppositely charged particle pairs tend to be aligned in the parallel direction for epsilon 1> 0 and in the perpendicular plane for epsilon 1< 0 with respect to the background director. For identically charged pairs the preferred directions for epsilon 1> 0 and epsilon 1< 0 are exchanged. We also examine competition between the charge-induced anchoring and the short-range anchoring. If the short-range anchoring is sufficiently strong, it can be effective in the vicinity of the surface, while the director orientation is governed by the long-range electrostatic interaction far from the surface.

Indoor organophosphate and polybrominated flame retardants in Tokyo.

UNLABELLED: In Japan, organophosphate and polybrominated flame retardants are used in building materials and electric appliances to protect them from fire hazards. In this study, to identify the emission sources of these flame retardants to indoor air, the migration rates (flux) of organophosphate and polybrominated flame retardants from building materials and electrical appliances to solid extraction disks that were placed in contact with the interior surfaces were measured. In addition to the migration test, indoor air and outdoor air concentrations of these flame retardants were investigated. With regard to building materials in a newly built house, triethylphosphate (TEP) and tributylphosphate (TBP) were detected in the wall and ceiling coverings, and tris(2-butoxyethyl)phosphate (TBEP) was detected in the wooden flooring cleaned with a floor polish agent. With regard to electrical appliances, triphenylphosphate (TPHP) was predominantly detected in computer monitors and tris(2-chloroethyl) phosphate (TCEP) in television (TV) sets, with the highest median levels. Among the polybrominated compounds, only 2,2',4,4'-tetrabromodiphenyl ether (BDE-47) was detected from a few old TV sets manufactured before 1995. In an indoor and outdoor air survey, nine organophosphates and nine polybrominated flame retardants were detected from indoor air. In outdoor air, only four organophosphate flame retardants were detected. The maximum level of indoor organophosphate compounds was 1260 ng/m(3) with tris(2-chloro-1-methylethyl) phosphate (TCPP), and that of polybrominated compounds was 29.5 ng/m(3) with hexabromocyclododecane (HBCD). Tetrabromobisphenol A (TBBPA) was not detected in this study, although it has the largest demand among flame retardants in Japan. The results of the migration test and the indoor air survey revealed that in indoor air, organophosphate compounds were more predominant than polybrominated compounds in Tokyo. PRACTICAL IMPLICATIONS: Polybrominated biphenyls (PBB) and polybrominated diphenyl ethers (PBDE) are commonly used as flame retardants in plastics. The use of these two compounds in electric appliances will be banned in 2007 by the EU Directives on waste electrical and electronic equipment (WEEE) and on the restriction of the use of certain hazardous substances (RoHS) in electrical and electronic equipment. In Japan, the use of PBB was banned and that of PBDE diminished in the early 1990s by the self-imposed controls of the Japanese Flame Retardants Conference (Akutu and Hori, 2004). In Japan, the predominantly used organic flame retardants were tetrabromobisphenol A and organophosphate compounds. Tetrabromobisphenol A has been reported to disrupt endocrine systems (Kitamura et al., 2005), and some organophosphate flame retardants were recently reported to have neurochemical hazardous effects. Furthermore, organophosphate compounds were suspected to cause endocrine-disrupting effects (Fang et al., 2003; Ohyama et al., 2005) or attention deficit hyperactivity disorder (ADHD) (Winrow et al., 2003). In this study, organophosphate and polybrominated flame retardants were surveyed in indoor environments in Tokyo.

Numerical simulation studies of the convective instability onset in a supercritical fluid.

Numerical simulation studies in 2D with the addition of noise are reported for the convection of a supercritical fluid, 3He , in a Rayleigh-Bénard cell. The noise addition is to accelerate the instability growth after starting the heat flow across the fluid, so as to bring simulations into better agreement with published experimental observations. Homogeneous temperature noise and spatial longitudinal periodic temperature variations in either top or bottom plates were programmed into the simulations. The second method was the most effective in speeding up the instability onset. For a small amplitude of the longitudinal perturbations, a semiquantitative agreement with the observations was obtained. The results are discussed in relation to predictions by El Khouri and Carlès.

Indoor air pollution by alkylphenols in Tokyo.

UNLABELLED: The purposes of this study were to develop an analytical method for airborne alkylphenols (APs) using the gas chromatography/mass spectrometry (GC/MS)-SIM method and to investigate airborne C4-C9 APs pollution in houses, offices, and outdoor points in Tokyo. In addition, to identify APs emission sources in a newly built house, migration tests of APs from interior surfaces to solid extraction disks were performed. Air samples were collected from houses, offices, and outdoor points. The recovery of APs from air samples was increased by addition of ascorbic acid to the filters. 4-tert-butylphenol (4-t-BP), 4-tert-octylphenol (4-t-OP), and 4-nonylphenol (4-NP) were detected in both indoor and outdoor air. Their concentrations and detection frequencies were higher in indoor air than outdoor air. The maximum levels of 4-t-BP, 4-t-OP, and 4-NP in indoor were 387, 45.7, and 680 ng/m(3), respectively. 4-t-BP and 4-NP were detected with high frequencies (more than 97%) in indoor air samples. The concentrations of 4-t-BP, 4-t-OP, and 4-NP in indoor air were significantly correlated with room temperature (P < 0.01), and tended to be higher in newly built houses. The migration rates of APs from the floor, wall, and ceiling in the newly built house were higher in polyvinyl chloride coverings than in other materials. Practical Implications Alkylphenol ethoxylate (APE) are widely used surfactants in both industrial and domestic detergents. When these detergents are released into the environment, APE are biodegraded to the smaller chain ethoxylates and alkylphenols (APs). There are many reports about APs in aquatic environments because of their estrogenic effect on wildlife. In building materials, APs are used as antioxidant for polymer resins such as wall or floor coverings. Although APs has a lower vapor pressure, these agents are released into indoor air from the surface of polymer resins. This study surveyed concentrations of airborne AP in indoor air and outdoor air in Tokyo. In addition, to identify APs emission sources in a newly built house, migration tests of APs from interior surfaces to solid extraction disks were performed. PRACTICAL IMPLICATIONS: Alkylphenol ethoxylate (APE) are widely used surfactants in both industrial and domestic detergents. When these detergents are released into the environment, APE are biodegraded to the smaller chain ethoxylates and alkylphenols (APs). There are many reports about APs in aquatic environments because of their estrogenic effect on wildlife. In building materials, APs are used as antioxidant for polymer resins such as wall or floor coverings. Although APs has a lower vapor pressure, these agents are released into indoor air from the surface of polymer resins. This study surveyed concentrations of airborne AP in indoor air and outdoor air in Tokyo. In addition, to identify APs emission sources in a newly built house, migration tests of APs from interior surfaces to solid extraction disks were performed.

Convection in a very compressible fluid: comparison of simulations with experiments.

The time profile DeltaT(t) of the temperature difference, measured across a very compressible fluid layer of supercritical 3He after the start of a heat flow, shows a damped oscillatory behavior before steady-state convection is reached. The results for DeltaT(t) obtained from numerical simulations and from laboratory experiments are compared over a temperature range where the compressibility varies by a factor of approximately 40. First the steady-state convective heat current j(conv) as a function of the Rayleigh number Ra is presented, and the agreement is found to be good. Second, the shape of the time profile and two characteristic times in the transient part of DeltaT(t) from simulations and experiments are compared, namely (1) t(osc), the oscillatory period, and (2) t(p), the time of the first peak after starting the heat flow. These times, scaled by the diffusive time tau(D) versus Ra, are presented. The agreement is good for t(osc)/tau(D), where the results collapse on a single curve showing a power-law behavior. The simulation hence confirms the universal scaling behavior found experimentally. However for t(p)/tau(D), where the experimental data also collapse on a single curve, the simulation results show systematic departures from such a behavior. A possible reason for some of the disagreements, both in the time profile and in t(p), is discussed.

Thermal plumes and convection in highly compressible fluids.

We present simple hydrodynamic equations of supercritical fluids close to the gas-liquid critical point. We numerically solve them to examine plume generation and convection under gravity. These results are in good agreement with the experiment [A. B. Kogan and H. Meyer, Phys. Rev. E 63, 056310 (2001)]. This Letter is a first study of transient behavior of convection, which is unique in compressible fluids due to the piston effect.

Phase transitions of binary alloys with elastic inhomogeneity.

In elastically inhomogeneous alloys, in which the shear modulus depends on the composition, coarsening in phase separation can be pinned even without quenched disorder. Here networks in the soft phase enclose the domains in the hard phase. Highly asymmetric strains in the two regions create large free energy barriers that prevent further coarsening. Thus a phase transition occurs between the one-phase and the glassy two-phase states. We numerically obtain the phase diagram and show that this transition is discontinuous at any composition. Therefore there is no critical point.

Detection of porcine circovirus from lesions of a pig with wasting disease in Japan.

A wasting disease characterized by progressive weight loss and dyspnea has been observed in weaning pigs on a farm in Yamagata Prefecture in 1998. Histopathologic findings in an affected pig were bronchointerstitial pneumonia and intracytoplasmic clusters of basophilic inclusions in macrophages of lymph nodes, which were similar to those in pigs with postweaning multisystemic wasting syndrome (PMWS) recently reported in North America and Europe. Porcine circovirus (PCV)-like particles were observed in bronchial lymph node of the pig by electron microscopy, and PCV antigens were detected in the lesions by immunohistochemical staining. PCV DNA was also detected in the lung and tonsil by PCR, and restriction fragment length polymorphism analysis of the PCR products with HinfI showed the same type of the PCV associated with PMWS (pmws PCV). Homology of nucleotide sequences between the PCR product and corresponding regions of published pmws PCV genomes was very high. These results indicated that virus detected in this study was pmws PCV. To our knowledge, this is the first report on the presence of pmws PCV in Japan.

Tracheal reconstruction by esophageal interposition: an experimental study.

The purpose of this study was to assess the possibility of reconstructing a circumferential tracheal defect with autogenous esophagus. In 6 mongrel dogs, a circumferential defect involving seven rings of the cervical trachea was reconstructed by interposing pedicled esophagus. A silicone T tube was used as a stent. The vertical limb of the T tube (usually referred to as the horizontal limb when used in humans) was cut shorter after the cervical wound had healed well, and it eventually was buried subcutaneously. Two dogs died 36 days after operation, and 1 died 28 weeks after operation. In no dog was the cause of death related to the operation or to a respiratory tract complication. Two dogs were put to death 4 weeks and 32 weeks after operation. They were well until then, and all the anastomoses between the trachea and the esophagus had healed fully without formation of granulation tissue. One dog is alive and well 14 months after operation. The vertical limb of the T tube retracted into the subcutaneous space, and there is no open cervical wound. Esophageal interposition might be a feasible technique for tracheal replacement in select groups of patients.