Effect of growth conditions of Lactobacillus gasseri OLL2809 on the immunostimulatory activity for production of interleukin-12 (p70) by murine splenocytes

Lactobacillus gasseri OLL2809, a probiotic lactic acid bacterium, strongly stimulates interleukin (IL)-12 (p70) production by murine splenocytes; therefore, it is expected to ameliorate allergic diseases. Although many studies have investigated characteristics of the immunostimulatory activity of probiotics, little is known about how bacterial growth conditions affect the activity. In this study, we investigated the effects of the growth conditions of L. gasseri OLL2809 on the stimulation of IL-12 (p70) production. L. gasseri OLL2809 was grown under various culture conditions including different cultivation periods, media, and culture pH, and IL-12 (p70) production by murine splenocytes stimulated with these bacterial cells was determined. The results revealed that IL-12 (p70) production (i) increased with the growth of the bacterial cells and was higher in stationary-phase cells than in logarithmic-phase cells; (ii) it was higher in the cells grown in acidic pH; and (iii) it decreased when the cells were incubated in a buffer at neutral pH prior to heat treatment. These observations indicated that stimulation of IL-12 (p70) production is affected by culture medium pH. In addition, the observations of a difference in the stimulation of IL-12 (p70) production by L. gasseri OLL2809 grown under various conditions are consistent with the characteristics of autolysis. Therefore, it was deduced that the integrity of the bacterial cells is necessary for the stimulatory effect on IL-12 (p70) production and that acidic pH and heat treatment contributed to the stimulation by inhibiting the activity of autolysins indigenous to the bacteria. Our result suggests that cultivation until the stationary phase under acidic pH is required for the effective production of probiotics with immunostimulatory activity.     

Electrical switching of the vortex core in a magnetic disk

A magnetic vortex is a curling magnetic structure realized in a ferromagnetic disk, which is a promising candidate for a memory cell for future non-volatile data-storage devices. Thus, an understanding of the stability and dynamical behaviour of the magnetic vortex is a major requirement for developing magnetic data-storage technology. Since the publication of experimental proof for the existence of a nanometre-scale core with out-of-plane magnetization in a magnetic vortex, the dynamics of vortices have been investigated intensively. However, a way to electrically control the core magnetization, which is a key for constructing a vortex-core memory, has been lacking. Here, we demonstrate the electrical switching of the core magnetization by using the current-driven resonant dynamics of the vortex; the core switching is triggered by a strong dynamic field that is produced locally by a rotational core motion at a high speed of several hundred metres per second. Efficient switching of the vortex core without magnetic-field application is achieved owing to resonance. This opens up the potentiality of a simple magnetic disk as a building block for spintronic devices such as a memory cell where the bit data is stored as the direction of the nanometre-scale core magnetization.     

Four-point probe resistance measurements using PtIr-coated carbon nanotube tips

We performed four-terminal conductivity measurements on a CoSi2 nanowire (NW) at room temperature by using PtIr-coated carbon nanotube (CNT) tips in a four-tip scanning tunneling microscope. The physical stability and high aspect ratio of the CNT tips made it possible to reduce the probe spacing down to ca. 30 nm. The probe-spacing dependence of resistance showed diffusive transport even at 30 nm and no current leakage to the Si substrate.     

Comparing the Osteogenic Potential and Bone Regeneration Capacities of Dedifferentiated Fat Cells and Adipose-Derived Stem Cells In Vitro and In Vivo: Application of DFAT Cells Isolated by a Mesh Method

Background: We investigated and compared the osteogenic potential and bone regeneration capacities of dedifferentiated fat cells (DFAT cells) and adipose-derived stem cells (ASCs). Method: We isolated DFAT cells and ASCs from GFP mice. DFAT cells were established by a new culture method using a mesh culture instead of a ceiling culture. The isolated DFAT cells and ASCs were incubated in osteogenic medium, then alizarin red staining, alkaline phosphatase (ALP) assays, and RT-PCR (for RUNX2, osteopontin, DLX5, osterix, and osteocalcin) were performed to evaluate the osteoblastic differentiation ability of both cell types in vitro. In vivo, the DFAT cells and ASCs were incubated in osteogenic medium for four weeks and seeded on collagen composite scaffolds, then implanted subcutaneously into the backs of mice. We then performed hematoxylin and eosin staining and immunostaining for GFP and osteocalcin. Results: The alizarin red-stained areas in DFAT cells showed weak calcification ability at two weeks, but high calcification ability at three weeks, similar to ASCs. The ALP levels of ASCs increased earlier than in DFAT cells and showed a significant difference (p < 0.05) at 6 and 9 days. The ALP levels of DFATs were higher than those of ASCs after 12 days. The expression levels of osteoblast marker genes (osterix and osteocalcin) of DFAT cells and ASCs were higher after osteogenic differentiation culture. Conclusion: DFAT cells are easily isolated from a small amount of adipose tissue and are readily expanded with high purity; thus, DFAT cells are applicable to many tissue-engineering strategies and cell-based therapies.     

Flow structure and heat transfer mechanism of laminar vortex rolls driven by gravity currents between parallel plates of nonuniform wall temperature

Laminar vortex rolls which occur between horizontal parallel plates, of which the temperature is not uniform in the lateral direction normal to the forced main flow, are investigated by means of a flow-visualization technique and a 3D steady-state numerical calculation. It was found that the vortex rolls were driven by the horizontal gravity currents, which were caused by density difference in the lateral direction, and that the generation mechanism of the vortex rolls could be classified into four types. The effects of vortex rolls on the local Nusselt number were also investigated. © 1999 Scripta Technica, Heat Trans Asian Res, 28(7): 606–619, 1999     

Accumulation of paralytic shellfish poison (PSP) and biotransformation of its components in oysters, Crassostrea gigas, fed with the toxic dinoflagellate Alexandrium tamarense

As a part of our studies on the mechanism of uptake of paralytic shellfish poison (PSP) and the kinetics of its accumulation in bivalves, oysters Crassostrea gigas were experimentally contaminated with PSP by being fed with the toxic dinoflagellate Alexandrium tamarense for 2, 4, 6, 8 and 10 days. Temporal variations in the PSP contents and their profiles in oysters during the feeding experiment were monitored by high-performance liquid chromatography (HPLC) and the toxin profile of the oysters was compared with that of A. tamarense. Toxins excreted from the infested oysters into the seawater for 2 and 10 days were recovered and analyzed by HPLC. PSP toxicity rapidly appeared in the tissues of oysters and their toxicity levels reached 0.6 (0.3), 2.2 (1.1), 1.0 (0.5), 3.4 (1.6) and 1.1 (0.5) MU/g (nmol/g) shucked meat at 2, 4, 6, 8 and 10 days, respectively. The accumulation rates of toxin, calculated from the total amount (nmol) of toxins expressed by the total cell number fed during the exposure period and the toxicity of the oysters, were 14.1, 18.7, 5.1, 14.9 and 3.2% for 2, 4, 6, 8 and 10 days. During feeding experiments, the toxin profile of oysters changed substantially, showing marked differences from the proportions found in the toxigenic dinoflagellate used as food. The toxin components in this strain existed almost exclusively as beta-epimers, which accounted for 66.3 mol% of the total. This contrasts with the case of the oysters, where the beta-epimers represented 24.8, 29.8, 25.1, 27.3 and 25.2 mol% of the total at 2, 4, 6, 8 and 10 days, respectively. The amount of gonyautoxin-1 (GTX1) accumulated in oysters increased linearly and slowly for 8 days and the maximum content of GTX1 reached 51.3 mol%. The composition of GTX group compounds recovered from the seawater in which the oysters had been reared was a little different from that within the oyster tissues.     

Precise metabolic flux analysis of coryneform bacteria by gas chromatography-mass spectrometry and verification by nuclear magnetic resonance

Precise metabolic flux analysis (MFA) by gas chromatography-mass spectrometry (GC-MS) and computer calculation was performed, and the consistency of the estimated results was verified by independently performed nuclear magnetic resonance (NMR) analysis. The precise estimation of flux by the integration method of the mass isotopomer signal, defined as the coefficient of variance (CV) of multiple determination, was investigated, and the results estimated using different data sets with the same magnitude of error were confirmed. The CV of multiple determinations was sufficiently small to discuss and compare the fluxes of a metabolic pathway. The estimated fluxes using the GC-MS data were cross-validated with the NMR data that were independently measured and not used for MFA. The developed method was successfully applied to the MFA of the growth phase of two different glutamate-producing coryneform bacteria, Corynebacterium glutamicum and C. efficiens. The difference in the growth rate between these two bacterial species was discussed while considering the results of MFA, including forward and backward (exchange) fluxes.     

Observation of the anisotropy of the inelastic scattering of fast electrons accompanied by the K-shell ionization of a carbon nanotube

The angular distribution of inelastically scattered electrons following the carbon K-shell ionization of a carbon nanotube (CNT) has been studied by using an energy-filtered transmission electron microscope. The energy-filtered scattering patterns of the CNT, whose cylindrical axis is oriented in the perpendicular direction with respect to the incident-electron beam, show anisotropic angular distributions caused by the p symmetry in the pi* and sigma* unoccupied states into which a K electron jumps, and the cylindrical shape of the CNT. Corresponding inelastic scattering cross sections, differential in solid angle, have been calculated within the first Born approximation by considering that each carbon atom has three, mutually orthogonal, unoccupied p orbitals, which are oriented in directions perpendicular or parallel to the tube surface. The observed angular patterns agree well with theoretical results, which are linear combinations of the differential cross sections accompanied by the transitions to pi* and sigma* states. A series of the fittings of the experimental patterns at successive values of energy loss provides magnitudes of the pi* and sigma* components, which can be called partial electron energy-loss spectra accompanied by the transitions to the pi* and sigma* states.     

Expression of aldehyde dehydrogenase gene increases hydrogen production from low concentration of acetate by Rhodobacter sphaeroides

Rhodobacter sphaeroides RV (RV) is a hydrogen-producing bacterium exhibiting the highest yield of hydrogen production from organic acids such as lactate and acetate, which are the byproducts of hydrogen fermentation by hydrogen-producing anaerobic bacteria. Co-fermentation of the RV strain with anaerobic bacteria is an efficient method of hydrogen production. However, less than 21 mM acetate is produced by the anaerobic bacteria, which is too low for efficient hydrogen production by the RV strain; it requires approximately 75 mM acetate. In this study, 2 distinct isozymes of aldehyde dehydrogenase from Rhodospirillum rubrum were separately overexpressed in the RV strain. The recombinant RV strains that were designated as RVAD1 and RVAD2, exhibited 13-fold higher ALDH activities than the wild-type RV strain. Hydrogen yields of both of the recombinant strains were 1.4-fold higher than that of the RV strain in 21 mM acetate. In 43 mM acetate, the RVAD1 strain showed higher yield, though the RVAD2 strain showed lower yield as compared to that of the RV strain. In 64 mM acetate and all concentrations of lactate tested (21, 43 and 64 mM), the yields of the recombinant strains were lower than those of the RV strain. The intact (empty) expression plasmid increased the ALDH activity and had little effect on the hydrogen production in acetate, however, it decreased the production in lactate. At the beginning of the fermentation process, when very little hydrogen had been produced, the recombinant strains expressing the ALDH gene consumed smaller amounts of acetate compared to the wild-type strain. We have discussed the effects of ALDH on hydrogen production in this report.     

Differentiation of animal fats from different origins: use of polymorphic features detected by Raman spectroscopy

Food safety requires the development of reliable techniques that ensure the origin of animal fats. In the present work, we try to verify the efficacy of using the polymorphic features of fats for discriminating animal-fat origins. We use Raman spectroscopy to collect the structural information of fat crystals. It is shown that a single Raman band at 1417 cm(-1) successfully differentiates pork fats from beef fats. This band is known to be characteristic of the β'-polymorph of fats. Pork fats show this band because they contain the β'-polymorph after rapid cooling to 0 °C. In beef-pork-fat mixtures, this band is not detected even in the presence of 50% pork fat; an addition of beef fat to pork fat is likely to produce a mixed fat with a completely different polymorphic behavior. This method seems to have the potential to detect beef products contaminated with pork-adipose tissue.