Production of vitamin B12 in genetically engineered Propionibacterium freudenreichii

Since the chemical synthesis of vitamin B12 requires more than 70 steps, the production of vitamin B12 has been achieved by microorganism fermentation with additional brief chemical modifications. In an effort to increase the productivity of vitamin B12, we tried to express 10 genes belonging to the hem, cob and cbi gene families involved in the synthesis of vitamin B12 in Propionibacterium freudenreichii, which is a known producer of vitamin B12. In a recombinant P. freudenreichii clone that harbored the expression vector containing a cobA, cbiLF, or cbiEGH, we obtained an increase in vitamin B12 production of 1.7-, 1.9-, and 1.5-fold higher, respectively, than that in the microorganism without any cloned genes in the expression vector pPK705. The cobU and cobS genes caused a slight increase in the production of vitamin B12. Furthermore, we achieved multigene expression in P. freudenreichii. In a recombinant P. freudenreichii clone that harbored an exogenous gene, hemA, from Rhodobacter sphaeroides and endogenous hemB and cobA genes, we successfully achieved the production of about 1.7 mg/l vitamin B12, 2.2-fold higher than that produced by P. freudenreichii harboring pPK705.     

Activated carbon adsorption of trichloroethylene (TCE) vapor stripped from TCE-contaminated water

Ground water contaminated with trichloroethylene (TCE) used in electronic, electric, dry cleaning and the like industries is often treated by air-stripping. In this treatment process, TCE in its vapor form is stripped from ground water by air stream and sometimes emitted into the atmosphere without any additional treatments. Activated carbon adsorption is one of the practical and useful processes for recovering the TCE vapor from the exhaust air stream. However, adsorption of the TCE vapor from the stripping air stream onto activated carbons is not so simple as that from dry air, since in the exhaust air stream the TCE vapor coexists with water vapor with relatively high concentrations. The understanding of the adsorption characteristics of the TCE vapor to be adsorbed on activated carbon in the water vapor-coexisting system is essential for successfully designing and operating the TCE recovery process. In this work, the adsorption equilibrium relations of the TCE vapor adsorption on activated carbons were elucidated as a function of various relative humidity. Activated carbon fibers (ACFs) were used as model activated carbon. The adsorption equilibrium relations were studied by the column adsorption method. The adsorption isotherms of TCE vapor adsorbed on sample ACFs were successfully correlated by the Dubinin-Radushkevich equation for both cases with and without coexistent water vapor. No effects of coexistent water vapor were found on the limiting adsorption volume. However, the adsorption characteristic energy was significantly reduced by the coexistence of water vapor and its reduction was successfully correlated with the equilibrium amount of water vapor adsorbed under the dynamic condition.     

Radiation doses to neonates during X ray computed tomography examinations

As the survival rate of newborns has increased, the number of X ray computed tomography (CT) examinations performed on neonates has been increasing. The exposure doses from CT examinations are known to be higher than those from conventional radiography. Although radiation sensitivity of neonates is higher than that of adults, there are few reports on dose estimates of neonates in CT examinations. Four cylindrical phantoms and one neonatal phantom have been developed to estimate doses to neonates during CT examinations. Using these phantoms and glass dosemeters, absorbed doses were measured. Estimated exposure doses to neonates were higher than those to adults, and our results suggest a need to optimise carefully CT examinations in newborns.     

Ultrasonic techniques for imaging and measurements in molten aluminum

In order to achieve net shape forming, processing of aluminum (Al) in the molten state is often necessary. However, few sensors and techniques have been reported in the literature due to difficulties associated with molten Al, such as high temperature, corrosiveness, and opaqueness. In this paper, development of ultrasonic techniques for imaging and measurements in molten Al using buffer rods operated at 10 MHz is presented. The probing end of the buffer rod, having a flat surface or an ultrasonic lens, was immersed into molten Al while the other end with an ultrasonic transducer was air-cooled to room temperature. An ultrasonic image of a character "N", engraved on a stainless steel plate immersed in molten Al, and its corrosion have been observed at 780 degrees C using the focused probe in ultrasonic pulse-echo mode. Because cleanliness of molten Al is crucial for part manufacturing and recycling in Al processing, inclusion detection experiments also were carried out using the nonfocused probe in pitch-catch and pulse-echo modes. Backscattered ultrasonic signals from manually added silicon carbide particles, with an average diameter of 50 microm, in molten Al have been successfully observed at 780 degrees C. For optimal image quality, the spatial resolution of the focused probe was crucial, and the high signal-to-noise ratio of the nonfocused probe was the prime factor responsible for the inclusion detection sensitivity using backscattered ultrasonic signals. In addition, it was found that ultrasound could provide an alternative method for evaluating the degree of wetting between a solid material and a molten metal. Our experimental results showed that there was no ultrasonic coupling at the interface between an alumina rod and molten Al up to 1000 degrees C; therefore, no wetting existed at this interface. Also because ultrasonic velocity in alumina is temperature dependent, this rod proved to be able to be used as an in-line temperature monitoring sensor under 1000 degrees C in molten Al.     

Strength of bond to bone and cytotoxicity of sintered bodies of hydroxyapatite/zirconia composite particles

Regarding sintered bodies of hydroxyapatite (HAP)/zirconia (PSZ) composite particles prepared by covering the surface of HAP particles with PSZ particles, their strength of the bond to bone after implanting them into an organism and their cytotoxicity were evaluated. Cytotoxicity tests were conducted by the colony formation method. Cytotoxicity was not observed in the sintered bodies of HAP/PSZ composite particles, or in sintered monolithic HAP body and sintered monolithic PSZ body. The strength of bond between a sintered body and a bone was evaluated by measuring the shear strength at the interface between them after a fixed period following implantation of a sintered body into a rabbit femur. In all cases of the sintered monolithic HAP body, and the sintered bodies of HAP/PSZ composite particles and monolithic PSZ body, when the implantation period increased, the shear strength of the sintered body/bone interface tended to increase. In particular, this tendency was very high in the case of sintered bodies of HAP/PSZ with weight ratios of 1/1.0 and 1/1.5, the shear strength for each reaching 20 MPa 12 months after implantation.     

Synthetic potential of molluscan sulfatases for the library synthesis of regioselectively O-sulfonated D-galacto-sugars

The substrate specificities of three molluscan sulfatases (E.C. 3.1.6.1; snail, abalone, and limpet origins) were investigated with assorted p-nitrophenyl (pNP) di-O-sulfonated beta-D-galactopyranosides and beta-lactosides [3,6-SO(3) Gal (1), 3',6'-SO(3) Lac (2), 4, 6SO(3) Gal (3), 2,6-SO(3) Gal (4), 3,4-SO(3) Gal (5), and 3,6-SO(3) GalNAc (6); Ac, acetyl; Gal, galactose; Lac, lactose] together with mono-O-sulfonated beta-D-galactopyranoside [pNP 3SO(3)-Gal (7)] and tri-O-sulfonated alpha-D-galactopyranoside [2,3,6-SO(3)-alpha-Gal (11)]. Some notable differences between the substrate specificity of the three sulfatases were disclosed; snail sulfatase hydrolyzed the 3O- and 2O-sulfo groups of 1 and 4, respectively, to afford 6SO(3) Gal (9) in high yields, while the abalone enzyme did not act on 4. Only the limpet enzyme could cleave the 3O-sulfo groups of 7 to give pNP beta-galactoside. In contrast, every enzyme could utilize 11 as a good substrate to afford a mixture of 6SO(3)-alpha-Gal (13) and 2,6-SO(3) alpha-Gal (12). None of the enzymes could cleave the O-sulfo groups of 5 and 6, which indicates that a primary 6O-sulfo group tends to promote the enzymatic hydrolysis of O-sulfo groups at the secondary positions.     

Budding of fowlpox and pigeonpox viruses at the surface of infected cells

Chick embryo fibroblasts and chorioallantoic membranes infected with fowlpox virus (FWPV) or pigeonpox virus (PPV) were examined by transmission and scanning electron microscopy. Immature virus particles were observed in finely granular areas, i.e. virus factories, of the cytoplasm. These particles had various forms depending on their stages of development. Many tubular structures were also seen in these regions. Mature virus particles with ellipsoidal or brick-shaped forms enclosing electron-dense cores were detected throughout the cytoplasm. Notably, there was a high frequency of virus budding at the cell surface, but only occasional virus wrapping in the cytoplasm. Another remarkable feature of the infected cells was accumulation of many virions just beneath the plasma membrane, indicating that this phenomenon is closely related to virus budding. Based on the observed frequency of budding, this mechanism seems to be the predominant way for FWPV and PPV to exit the cell.     

Terahertz radiation from a shallow incidence-angle InAs emitter in a magnetic field irradiated with femtosecond laser pulses

The optimized incidence angle and magnetic field direction geometry of an InAs terahertz radiation emitter irradiated with femtosecond laser pulses in a magnetic field is reported. The optimum geometric layout is the magnetic field direction parallel to the semiconductor surface and at an incidence angle that is slightly larger than the Brewster angle. Additionally, we also observed a center frequency shift of terahertz radiation spectrum by changing the incidence angle of the excitation laser.     

Vibrational properties of wood along the grain

The dynamic Young's modulus (E_L) and loss tangent (tan _L ) along the grain, dynamic shear modulus (G_L) and loss tangent (tan _S) in the vertical section, and density () of a hundred spruce wood specimens used for the soundboards of musical instruments were determined. The relative acoustic conversion efficiency ( ) and a ratio reflecting the anisotropy of wood (, (E_L/G_L)(tan _S/tan _L)) were defined in order to evaluate the acoustic quality of wood along the grain. There was a positive correlation between and , and the variation in was larger than that in . It seemed logical to evaluate the acoustic quality of spruce wood by a measure of . By using a cell wall model, those acoustic factors were expressed with the physical properties of the cell wall constituents. This model predicted that the essential requirement for an excellent soundboard is smaller fibril angle of the cell wall, which yields higher and higher . On the other hand, the effects of chemical treatments on the and of wood were clarified experimentally and analyzed theoretically. It was suggested that the and of wood cannot be improved at the same time by chemical treatment.     

Power generation of the touching Cu/Bi-Te/Cu composites under the periodically alternating temperature gradients

The thermo-emf ΔV of the touching p- and n-type Cu/Bi-Te/Cu composites with different thicknesses of t _Bi-Te and t _Cu was measured as a function of time by alternating the temperature difference ΔT at periods of T = 20, 60, 120, 240 and ∞ sec, where t _Bi-Te was varied from 0.1 to 2.0 mm and t _Cu from 0 to 4.0 mm. As a result, ΔV changes significantly with t _Bi-Te, t _Cu and T. The effective thermo-emf ΔV _eff increases significantly with an increase of 1/T and exhibited a local maximum at 1/T = 1/240 s⁻¹. The resultant | α | and the effective temperature difference ΔT _eff were increased significantly by optimizing t _Bi-Te and t _Cu at 1/T = 1/240 s⁻¹. The power generation ΔW _eff (= ΔV _eff²/4R _calc) estimated using the measured ΔV _eff and calculated R _calc also exhibited a local maximum at 1/240 s⁻¹ for an optimum combination of t _Bi-Te = 0.1 mm and t _Cu = 2.0 mm, so that the maxima ΔW _eff at 1/T = 1/240 s⁻¹ for the p- and n-type composites were 2.28 and 2.92 times higher than those obtained at 1/T = 0 s⁻¹. This significant increase in ΔW _eff is owing to both the increase in ΔT _eff and the increase in ZT due to the increase in |α|. The power generation was thus found to be enhanced significantly by imposing the alternating temperature gradients on touching Cu/Bi-Te/Cu composites.