Hydrogen-rich pure water prevents superoxide formation in brain slices of vitamin C-depleted SMP30/GNL knockout mice

Hydrogen is an established anti-oxidant that prevents acute oxidative stress. To clarify the mechanism of hydrogen’s effect in the brain, we administered hydrogen-rich pure water (H₂) to senescence marker protein-30 (SMP30)/gluconolactonase (GNL) knockout (KO) mice, which cannot synthesize vitamin C (VC), also a well-known anti-oxidant. These KO mice were divided into three groups; recipients of H₂, VC, or pure water (H₂O), administered for 33 days. VC levels in H₂ and H₂O groups were <6% of those in the VC group. Subsequently, superoxide formation during hypoxia-reoxygenation treatment of brain slices from these groups was estimated by a real-time biography imaging system, which models living brain tissues, with Lucigenin used as chemiluminescence probe for superoxide. A significant 27.2% less superoxide formed in the H₂ group subjected to ischemia-reperfusion than in the H₂O group. Thus hydrogen-rich pure water acts as an anti-oxidant in the brain slices and prevents superoxide formation.     

Isolation and Characterization of a Marine Cyclohexylacetate-Degrading Bacterium Lutimaribacter litoralis sp. nov., and Reclassification of Oceanicola pacificus as Lutimaribacter pacificus comb. nov.

A novel aerobic, Gram-negative, non-motile, pleomorphic, and rod-shaped bacterium designated KU5D5^T was isolated from seawater that was obtained from the coastal region of the Goto Islands, Japan, on the basis of its ability to utilize cyclohexylacetate as the sole source of carbon and energy. Strain KU5D5^T grew at pH 6.0–8.0 and 10–35 °C in the presence of 1.0–5.0 % (w/v) NaCl. Analysis of the 16S rRNA gene sequence revealed that this strain was affiliated to the family Rhodobacteraceae in the class Alphaproteobacteria and was related most closely to Lutimaribacter saemankumensis (96.6 % similarity) and Oceanicola pacificus (96.6 %). The predominant respiratory lipoquinone was ubiquinone-10 and the major cellular fatty acids were C_18:1 ω7c (66.7 %), C_16:0 (7.7 %), C_12:1 3-OH (6.1 %), and C_17:0 (6.1 %). The DNA G+C content was 58.9 mol %. On the basis of physiological, chemotaxonomic, and phylogenetic data, strain KU5D5^T is suggested to represent a novel species of the genus Lutimaribacter, for which the name Lutimaribacter litoralis sp. nov. is proposed. It is also proposed that O. pacificus should be transferred to the genus Lutimaribacter as Lutimaribacter pacificus comb. nov. The type strain of L. litoralis is KU5D5^T (=JCM 17792^T = KCTC 23660^T) and the type strain of L. pacificus is W11-2B^T (=CCTCC AB 208224^T = LMG 24619^T = MCCC 1A01034^T).     

Determination of D- and L-enantiomers of methionine and [2H3]methionine in plasma by gas chromatography-mass spectrometry

A method for the stereoselective determination of D- and L-enantiomers of both methionine and [(2)H3]methionine in rat plasma was developed using gas chromatography-mass spectrometry with selected-ion monitoring (GC-MS-SIM). DL-[(2)H7]Methionine was used as analytical internal standard to account for losses associated with the extraction, derivatization and chromatography. The amino acids were purified by cation-exchange chromatography using BondElut SCX cartridge and derivatized with HCl in methanol to form methyl ester followed by subsequent N-acylation with optically active (+)-alpha-methoxy-alpha-trifluoromethylphenylacetyl chloride to form diastereomeric amide. Quantification was performed by SIM of the molecular-related ions of the diastereomers on the chemical ionization mode. Endogenous L-methionine concentrations in 50 microl of rat plasma were measured with relative intra- and inter-day precision of 4.0 and 6.3%, respectively. The intra- and inter-day reproducibility in the amounts of D- and L-[(2)H3]methionine determined were in good agreement with actual amount added.     

First Isolation of Cryptococcus magnus from a Cat

A 6-month-old male Japanese domestic cat with otitis externa due to Aspergillus fumigatus was treated with antifungal agents for 25 days and appeared to be cured. Many yeast colonies however developed from the ear canal samples on Sabouraud's dextrose agar at 27 degrees C for 5 days, instead of A. fumigatus. This yeast colony was cream-colored and slim in texture with smooth and highly glossy surface after 5-day incubation on Sabouraud's dextrose agar at 27 degrees C. The isolate was identified as Cryptococcus magnus by mycological analysis and 28S ribosomal analysis.     

The application of phenylmethanethiol and benzenethiol derivatives as odorless organosulfur reagents in the synthesis of thiosugars and thioglycosides

p-octyloxyphenylmethanethiol and p-dodecylbenzenethiol were prepared as new odorless organosulfur reagents. Thiosugars and thioglycosides were synthesized using these reagents without encountering any malodorous procedures.     

Gene transfection activities of amphiphilic steroid-polyamine conjugates

The design and evaluation of a novel potent class of DNA delivery agents based on steroid-polyamine conjugates bearing a flexible linker are reported. The hydrophobic regions are based on steroids, i.e. chlolestane and lithocholic acid motifs. The linker, which couples a hydrophobic steroid and a hydrophilic polyamine, in this study can be regarded as a two-atom extension of the conventional carbamate linker. We found that the gene transfection activity of the steroid-polyamine conjugates is influenced by the polyamine chain length and steroid structure. Molecular modeling of the relevant amphiphilic molecules revealed low-energy structures in which the polyamine chains are folded rather than stretched. This work suggests a significant effect of space-filling, i.e. the shape and orientation of the hydrophilic and hydrophobic regions, upon the efficiency of gene transfection.     

Assessment of poliovirus eradication in Japan: genomic analysis of polioviruses isolated from river water and sewage in toyama prefecture

Seventy-eight poliovirus strains isolated from river water and sewage in Toyama Prefecture, Japan, during 1993 to 1995 were characterized by the PCR-restriction fragment length polymorphism (RFLP) method and by partially sequencing the VP3 and VP1 regions of the viral genome. Of these isolates, 36 were identified as Sabin vaccine strains, and 42 were identified as vaccine variant strains that had less than 1.4% nucleotide divergence from the Sabin strains, including 7 isolates with patterns different from those of Sabin strains as determined by PCR-RFLP analysis. These findings suggest that wild-type poliovirus was not circulating in Toyama Prefecture.     

Repetition of the classical Boysen-Jensen and Nielsen's experiment on phototropism of oat coleoptiles

The classical experiment of phototropic response as reported by Boysen-Jensen and Nielsen (1926), which supports the Cholodny-Went theory, was repeated in detail. In the original experiment, etiolated oat (Avena sativa L. cv. Victory) coleoptiles with mica inserted into their tip only showed a positive response when the mica was placed parallel toward the light source and not if it was inserted perpendicularly. On the contrary, we found a positive response irrespective of whether the mica was inserted parallel or perpendicularly to the light source. Damage owing to rude splitting severely reduced the response upon perpendicular insertion. These results invalidate the Boysen-Jensen and Nielsen's experiment as a support of the Cholodny-Went theory and lend support to the Bruinsma-Hasegawa theory ascribing phototropism to the local light-induced accumulation of growth inhibitors against a background of even auxin distribution, the diffusion of auxin being unaffected.