Overexpression of 1-Deoxy-D-xylulose-5-phosphate reductoisomerase gene in chloroplast contributes to increment of isoprenoid production

Plants synthesize a large number of isoprenoid compounds that are of industrial, nutritional and medicinal importance. 1-Deoxy-D-xylulose reductoisomerase (DXR) catalyzes the first committed step of plastidial isoprenoid-precursor biosynthesis. In the present study, we generated transplastomic tobacco plants that overproduced DXR from Synechosystis sp. strain PCC6803. The transformants showed increase in the content of various isoprenoids such as chlorophyll a, beta-carotene, lutein, antheraxanthin, solanesol and beta-sitosterol, indicating that the DXR reaction is one of the key steps controlling isoprenoid level in tobacco leaves. A qualitative change in isoprenoid composition was also observed. The growth phenotype of the transplastomic plants was similar to that of wild-type plants. These results showed that plastid metabolic engineering is useful in manipulating the yield of isoprenoids in plants.     

Ultrahighly sensitive in situ metabolomic imaging for visualizing spatiotemporal metabolic behaviors

A sensitive and simultaneous analytical technique for visualizing multiple endogenous molecules is now strongly required in biological science. Here, we show the applicability of a matrix-assisted laser desorption ionization mass spectrometry (MALDI-MS) system for getting chemically diverse metabolite profiles on a single-mammalian cell. This ultrahighly sensitive MALDI-MS technique enabled a spatially resolved detection of a broad range of metabolites including nucleotides, cofactors, phosphorylated sugars, amino acids, lipids, and carboxylic acids in normal mouse brain tissue with their unique distributions. Furthermore, a combination of MS imaging and metabolic pathway analysis of a rat transient middle cerebral artery occlusion model visualized a spatiotemporal behavior of metabolites in the central metabolic pathway regulated by an ischemia reperfusion. These findings highlight potential applications of an in situ metabolomic imaging technique to visualize spatiotemporal dynamics of the tissue metabolome, which will facilitate biological discovery in both preclinical and clinical settings.     

Synthesis and properties of polycyclic quinones condensed with 1,6-methano[10]annulene

Two types of polycyclic quinones condensed with 1,6-methano[10]annulenes as type A: 1,6-methanonaphtho[2,3-c][10]annulene-7,12-dione 5a, and type B: 1,6-methanonaphtho[2,3-c][10]annulene-5,14-dione 18, bis(1,6-methano[10]annuleno[3,4-b; 3,4-g])anthracene-10,21-dione 20, 1,6-methanoanthraceno[2,3-c][10]annulene-5,16-dione 22, 1,6-methanotetraceno[2,3-c][10]annulene-6,17-dione 23, and 1,6-methano phenanthreno[2,3-c][10]annulene-5,6-dione 24 have been synthesized. The acene derivative 6 corresponding to that of 5a was synthesized by the reduction of quinone 5a. The physical, spectral, and chemical properties of these new compounds have been investigated.     

Silica and titanium dioxide nanoparticles cause pregnancy complications in mice

The increasing use of nanomaterials has raised concerns about their potential risks to human health. Recent studies have shown that nanoparticles can cross the placenta barrier in pregnant mice and cause neurotoxicity in their offspring, but a more detailed understanding of the effects of nanoparticles on pregnant animals remains elusive. Here, we show that silica and titanium dioxide nanoparticles with diameters of 70 nm and 35 nm, respectively, can cause pregnancy complications when injected intravenously into pregnant mice. The silica and titanium dioxide nanoparticles were found in the placenta, fetal liver and fetal brain. Mice treated with these nanoparticles had smaller uteri and smaller fetuses than untreated controls. Fullerene molecules and larger (300 and 1,000 nm) silica particles did not induce these complications. These detrimental effects are linked to structural and functional abnormalities in the placenta on the maternal side, and are abolished when the surfaces of the silica nanoparticles are modified with carboxyl and amine groups.     

Relationship between Pectic Substances and Strand Separation of Cooked Spaghetti Squash--Part 2. Changes in Firmness,Histological Structure and Pectic Substances during Soaking in Pectin Extractants

The flesh of spaghetti squash separates into strands when cooked. The purpose of this paper is to investigate the cause of strand separation （during cooking） by soaking for 24 h at 35 ℃ in solutions with three kinds of pectin extractant. The changes in strand separation, firmness, histological structure and the pectin of flesh during soaking in 0.01 N HCI solution （pH 2.0）, 0.035 M ammonium oxalate solution （pH 4.0） or 2% sodium hexametaphosphate solution （pH 4.0） were investigated. When flesh was soaked in the HCI solution, the separation into strands and removal of calcium and magnesium were greater than that soaked in other pectin extractants. High methoxyl pectin was extracted by soaking in HC1 solution （pH 2.0） due to removal of polyvalent cations. This result shows that high methoxyl pectin glues strands together in the flesh of spaghetti squash. The shape of the cells which constituted strands was round; on the other hand, that of cells surrounded strands was elongated. When cooked in boiling water or soaked at pH 2.0, the shape of the former cells was maintained, but the latter cells, which contributed to adhesion between strands, broke down. Thus, the flesh separated into strands. When flesh was boiled for 15-30 min, pectin degraded and dissolved in the cooking solution; consequently, the flesh separated into strands and also the middle lamella of cell walls of strands separated. However, pectin remaining in strands maintained their crispness.     

Kinetic analysis of E. coli inactivation by high hydrostatic pressure with salts

Inactivation of E. coli by high hydrostatic pressure (250 to 400 MPa) with salts was investigated based on kinetic analysis. At concentrations from 0.074 to 0.145 M and from 0.240 to 0.290 M, both the absolute activation volumes and the preexponential factors were similar in KCl, NaCl, and LiCl solutions, suggesting that pressure inactivation is not salt-specific. On the other hand, in the intermediate salt-concentration range of 0.145 to 0.240 M, inactivation kinetics in the presence of the Na(+) and K(+) differed significantly from those in the presence of Li(+) (P < 0.05). In this concentration range, effect of salt stress and osmotic stress differed significantly from those in concentrations below 0.145 M or above 0.240 M. The cellular response to pressure varies with salt type and salt concentration. These novel findings provide important clues to distinguish between salt stress and osmotic stress in the inactivation of E. coli.     

Effects of exogenous mevalonic acid on sterol lipid classes in Larix kaempferi callus

In Japanese larch (Larix kaempferi (Lamb.) Carr.) calli, free sterol (FS), acylsterol (AS) and glycosylsterol, including the acylated type, were found in the proportion of 1.0:0.1:0.8. When the calli were cultured in the presence of 10 mM mevalonic acid (MVA), the content of AS, but not FS and glycosylsterol, was increased remarkably. The major component sterol in each sterol lipid class was usually sitosterol (more than 90%) with campesterol as a minor one. There were no differences on the sterol compositions between the calli cultured with or without MVA. When the calli cultured with 10 mM MVA for 6 weeks were transferred to the control medium without exogenous MVA, AS contents decreased to the level of the control calli. Thus, it was shown that sterol lipids, such as FS and glycosylsterols, with the structural functions was maintained in the constant content and the excess sterol biosynthesized from exogenous MVA was esterified to form AS for storage of sterol components.     

Cloning and nucleotide sequence of carbaryl hydrolase gene (cahA) from Arthrobacter sp. RC100

A carbaryl hydrolase gene (cahA) encoded on the plasmid pRC1 in Arthrobacter sp. RC100 was cloned and sequenced. The entire region of the deduced amino acid sequence was found to be homologous to that of an amidase family. Parts of the consensus sequences of the amidase gene have been identified in CahA from strain RC100. CahA was overexpressed in Escherichia coli JM109, and the enzyme was purified to homogeneity by protamine sulfate treatment, ammonium sulfate precipitation, and hydrophobic and anion-exchange chromatographies. The purified enzyme showed hydrolase activity toward 1-naphthylacetamide and isobutyramide but showed no activity toward 1-naphthylacetate. This is the first report of an amidase that is able to hydrolyze N-methylcarbamate pesticides.     

Enhanced bactericidal action and masking of allergen structure of soy protein by attachment of chitosan through maillard-type protein-polysaccharide conjugation

The soy protein-chitosan conjugate was formed by the Maillard reaction in dry state (relative humidity 65%) at 60 degrees C for 2 weeks to improve the functional properties. The antimicrobial activity of the Maillard-type soy protein-chitosan conjugates enhanced 2-3 times that of soy protein-chitosan mixture. The soy protein-chitosan conjugate showed excellent emulsifying property with the progress of Maillard-type conjugation. The allergenicity of soy protein was greatly decreased by the attachment of chitosan through Maillard reaction. The immonoblotting analysis with patient's sera revealed that soy protein-chitosan conjugate was more effective to mask the allergen structure of soy protein causing from 34 kDa-protein (Gly m Bd 30K) than soy protein-galactomannan conjugate. The Western blotting showed that allergen (34 kDa-protein) was completely masked by soy protein-chitosan conjugation, while it was not completely masked by soy protein-galactomannan conjugation.     

Effects of different triggering systems and external PEEP on trigger capability of the ventilator

OBJECTIVE: The triggering capability of both the pressure and flow triggering systems of the Servo 300 ventilator (Siemens-Elema, Sweden) was compared at various levels of positive end-expiratory pressure (PEEP), airway resistance (R(aw)), inspiratory effort and air leak, using a mechanical lung model. DESIGN: The ventilator was connected to a two bellows-in-series-type lung model with various mechanical properties. Lung compliance and chest wall compliance were 0.03 and 0.121/cmH2O, respectively. R(aw) was 5, 20 and 50 cmH2O/l/s. Respiratory rate was 15 breaths/min. To compare the triggering capability of both systems, the sensitivity of pressure and flow triggered pressure support ventilation (PSV) was adjusted to be equal by observing the triggering time at 0 cmH2O PEEP and 16 cmH2O of pressure support (PS) with no air leak. No auto-PEEP was developed. In the measurement of trigger delay, the PS level ranged from 16 to 22 cmH2O to attain a set tidal volume (V(T)) of 470 ml at a R(aw) of 5, 20 and 50 cmH2O/l/s. The PEEP level was then changed from 0, 5 and 10 cmH2O at a PS level of 17 cmH2O and R(aw) of 5 and 20 cmH2O/l/s, and the trigger delay was determined. The effect of various levels of air leak and inspiratory effort on triggering capability was also evaluated. Inspiratory effort during triggering delay was estimated by measurements of pressure differentials of airway pressure (Paw) and driving pressure in the diaphragm bellows (Pdriv) in both systems. MEASUREMENTS AND RESULTS: There were no significant differences in trigger delay between the two triggering systems at the various PEEP and R(aw) levels. At the matched sensitivity level, air leak decreased trigger delay in both systems, and additional PEEP caused auto-cycling. A low inspiratory drive increased trigger delay in the pressure sensing system, while trigger delay was not affected in the flow sensing system. The Paw and Pdriv differentials were lower in flow triggering than in pressure triggering. CONCLUSIONS: With respect to triggering delay, the triggering capabilities of the pressure and flow sensing systems were comparable with and without PEEP and/or high airway resistance at the same sensitivity level, unless low inspiratory drive and air leak were present. In terms of pressure differentials, the flow triggering system may require less inspiratory effort to trigger the ventilator than that of the pressure triggering system with a comparable triggering time. However, this difference may be extremely small.