Proteome analysis of cerebrospinal fluid in healthy beagles and canine encephalitis

We performed proteomics analysis of the cerebrospinal fluid (CSF) of healthy dogs and dogs with meningoencephalitis of unknown etiology (MUE). By comparing two-dimensional electrophoreses (2DE), an upregulated spot was found in MUE dogs. This protein was identified as a neuron-specific enolase (NSE) by analysis with MALDI-TOF mass spectrometry. In comparing dot blots using an antibody against NSE, the NSE levels in the CSF of MUE dogs was significantly higher than that of the controls. NSE is a diagnostic marker of neuroendocrine tumors, brain injury and spinal cord trauma in humans. It seems that the NSE concentration in the CSF is increased by cellular destruction in canine encephalitis. Though elevation of NSE may not be specific in canine encephalitis because the NSE level was increased in other CNS diseases, further study including measurement with serum is necessary.     

Ozone treatment of spent medium from Auricularia polytricha cultivation for enzymatic saccharification and subsequent ethanol production

We examined the enzymatic saccharification and ethanol fermentation of the spent media (SMs) from Auricularia polytricha cultivation using wood meals of Falcataria moluccana, Shorea sp., and Tectona grandis. Although the hydrolysis weight decrease and reducing sugar yield were higher in SM of F. moluccana, the ethanol yield was higher in SM of Shorea sp. Ozone treatment of SM further increased the hydrolysis weight decrease, reducing sugar, and ethanol yields in Shorea sp. These results indicate that SM of A. polytricha is a suitable biomass material to produce fermentable sugars for ethanol production, and that ozone treatment is a suitable method for increasing the ethanol yield.     

Contributions of the lipopolysaccharide outer core oligosaccharide region on the cell surface properties of Pseudomonas aeruginosa

We have characterized the cell surface properties of three mutant series of Pseudomonas aeruginosa that show various defects in their lipopolysaccharide (LPS) core region. The deepest rough mutants of each series used in this study lacked completely rhamnose and glucose, and contained only galactosamine and alanine as LPS outer core constituents. However, rough mutants other than the deepest rough mutants showed high cell surface hydrophobicity compared to the corresponding parental strains, the deepest rough mutants showed less hydrophobicity than other rough mutants. The reactivity of an anti-lipid A monoclonal antibody with the deepest rough mutants was markedly higher than that with other counterparts. The deepest rough mutants tended to be more susceptible to antibiotics, such as gentamicin and polymyxin B, than the corresponding parental strains and other rough mutants. The above evidence indicates that neutral sugar, namely rhamnose and glucose, residues of the LPS outer core region play a critical role in the cell surface properties of P. aeruginosa.     

Study on sheet material made from zephyr strands V: Properties of zephyr strand board and zephyr strand lumber using the veneer of fast-growing species such as poplar

Zephyr strand board (ZSB) and zephyr strand lumber (ZSL) were produced using zephyr made from poplar veneer to investigate the greater utilization of low-density poplar as a structural material. These materials were then compared to ordinary plywood, laminated veneer lumber (LVL) from poplar veneer, lauan plywood, and particleboard. The bending properties (moduli of rupture and elasticity) of ZSB proved superior to those of poplar plywood: and ZSL produced from poplar veneer zephyr had bending properties greater than ordinary LVL from poplar veneer. Apparently, the conversion of the poplar veneer into zephyr material had a positive effect on bending properties. Additionally, poplar ZSB had bending properties superior to those of lauan particleboard and equal to those of lauan plywood. The internal bond strength of poplar veneer ZSB was nearly two times greater than that of lauan particleboard.Parts of this report were presented at the international symposium on the utilization of fast-growing trees, Nanjing, China, October 1994. Report IV appeared inMokuzai Kogyo 49:599, 1994     

How to determine the appropriate mortality in experimental larval rearing?

Survival in larval rearing experiments is difficult to estimate due to accidental losses and periodic sampling. The number of sampled fish can be a large proportion of the stocked ones, making it difficult to calculate the overall survival rate and mortality coefficient as this is based on the initial number. Here, a new method of calculating survival is proposed using the mortality coefficient. When the initial stocking density and sampled and final numbers are known, and assuming that mortality coefficient is constant, the final number of fishes can be represented by the formula N _t = e^−mt (N ₀ − ΣN _Sne ^mdn), where t is rearing period (days), N ₀ indicates initial number, N _t indicates the survival number at t days of rearing, m is the natural mortality coefficient, N _Sn is the sampled number in the nth sampling, and dn is the rearing period until removal of the nth sample. The provisional mortality coefficient is calculated from initial and final stocking numbers. Then values for the natural mortality coefficient are substituted into the formula with successive approximation. The coefficient, which most closely approximates the actual survival, is determined as the best fit natural mortality coefficient. Examples of larval experiments are provided to demonstrate the method and show that survival is often underestimated using traditional methods.     

Temperature characteristics in seed germination and growth of <Emphasis Type="Italic">Zostera japonica</Emphasis> Ascherson &; Graebner from Ago Bay,Mie Prefecture,central Japan

The optimal water temperature in seed germination and the upper critical water temperature in seedling growth were determined for Zostera japonica collected from Ago Bay, Japan. The relationship between the seed germination rates and seed storage period (0, 30, and 60 days) at 0°C was also examined. The optimal water temperature in seed germination was in the range 15–20°C regardless of the storage period, in which germination rates were up to 14%. Seedlings, grown from seeds up to 10 cm in total length, were cultured for 1 week at various water temperatures to measure their relative growth rates. The optimal water temperature in early growth was in the range 20–25°C; relative growth rates ranged from 3.8 to 4.2%. Seedlings could survive up to a water temperature of 29°C, but most seedlings withered at 30 or 35°C. The optimal water temperatures for seed germination and seedling growth were related to the seasonal changes of water temperature in the sampling site. Although seedlings were hardly observed in Ago Bay in summer, Z. japonica might extend its distribution as far as where the summer water temperature is lower than 29°C.     

Preliminary study on homing,site fidelity,and diel movement of black rockfish <Emphasis Type="Italic">Sebastes inermis</Emphasis> measured by acoustic telemetry

Several species of fish exhibit strong homing ability and distinct site fidelity to their original habitats and spawning sites. We have studied the homing ability, site fidelity, and diel movement patterns of three nocturnal black rockfish Sebastes inermis. The patterns were studied by radio-linked acoustic positioning system (VRAP) and fixed monitoring receivers (VR2) in Maizuru Bay, Japan from June to July 2004. All tagged fish were displaced by 80–120 m, but they returned to their respective captured sites at dusk, regardless of the time of release. The fish tagged with a depth sensor transmitter displayed active vertical movement just after the release, and subsequently swam near the bottom depth around the captured site. After homing, they displayed small home ranges [average 815 ± 265 m² (SD)] around a single rocky area with one or two core areas (average 64 ± 30 m²). One core area was utilized during the daylight and dark periods, whereas the other was used primarily during the dark period. The depth-tagged fish primarily used the depth of the rock crevice during the daylight, and it spent its dark period between the surface and the deeper bottom depth.     

Mechanism of the rice hull‐induced germination of Monochoria vaginalis seeds in darkness

The seeds of Monochoria vaginalis require light to germinate. However, they are able to germinate in darkness in the presence of rice hulls. The mechanism of this phenomenon was investigated in this study, in which only the unsterilized seeds of Monochoria were induced to germinate by the presence of a rice hull extract. The rice hull extract was able to be replaced by either a mixture of amino acids and phosphate or a yeast extract. The culture media in which germination was observed always showed turbidity, indicating the propagation of bacteria, which were identified as Bacillus sp., Pedobacter sp., Pantoea sp. and Sphingomonas sp. Treatment with the individual bacterial species or combinations of these bacteria induced Monochoria seed germination when added to the media containing the rice hull extract. It was concluded that the rice hull extract accelerates the propagation of bacteria, which in turn digest the seed coat of Monochoria, facilitating its germination.     

Establishment of testicular and ovarian cell lines from Honmoroko (Gnathopogon caerulescens)

We succeeded to establish cell lines from endemic fish species Honmoroko Gnathopogon caerulescens, which inhabits Lake Biwa, the third oldest lake in the world. Two cell lines designated as RMT1 and RMO1 were established from testis and ovary of G. caerulescens, respectively. These cell lines were initially cultured in Leibovitz’s L-15 medium supplemented with fetal bovine serum (FBS), fish embryo extract, epidermal growth factor, and basic fibroblast growth factor. Further addition of forskolin and β-mercaptoethanol was required to establish and maintain these cell lines for more than 60 passages. RMT1 and RMO1 cells showed fibroblast- and epithelial-like morphology, respectively. From immunocytochemical staining and gene expression patterns, RMT1 cells showed a characteristic of testicular Sertoli cells and RMO1 cells did that of ovarian theca cells. Both RMT1 and RMO1 cells multiplied well in the medium supplemented with 10 % FBS at 28 °C and their minimum population doubling times were 24.4 and 28.8 h, respectively. At the 45th passage, most of the RMT1 and RMO1 cells had a hyperploid set of chromosomes (67.3 and 96.1 %, respectively). Cells with normal diploid chromosome set were not observed. RMT1 cells were transfected with an enhanced green fluorescent protein (EGFP) expression vector and human elongation factor 1 α promoter worked efficiently to express EGFP. In addition, EGFP-expressing cell lines were also established, suggesting that the cell lines could be utilized as an in vitro monitor system (biosensor) for the evaluation of endocrine disruptors which might affect gonadal function.     

Relationship between <Superscript>137</Superscript>Cs concentration and potassium content in stem wood of Japanese cedar (<Emphasis Type="Italic">Cryptomeria japonica</Emphasis>)

To utilize forest resources in areas affected by fallout from the Fukushima Daiichi Nuclear Power Plant accident, it is important to understand the mechanisms of ¹³⁷Cs movement through the stem wood of contaminated trees. Understanding the mechanism of absorption and migration of ¹³⁷Cs to stem wood is necessary for clues to the future prediction of the transition of ¹³⁷Cs to xylem. In the present study, radial variations in ¹³⁷Cs concentration were investigated in Japanese cedar (Cryptomeria japonica D. Don) trees collected 1 year and 10 months after the accident. Additionally, the relationship between ¹³⁷Cs concentration and potassium (K) content was established. Trees with a higher moisture content and lower lightness value in heartwood tended to have a higher ¹³⁷Cs concentration in the heartwood. In these trees, ¹³⁷Cs concentration peaked at the heartwood–sapwood boundary and gradually decreased toward the pith. By contrast, K content within the heartwood remained nearly constant along the radial direction. The heartwood-to-sapwood ratio of ¹³⁷Cs concentration was significantly positively correlated with that of K content. Based on these results, we suggest that ¹³⁷Cs movement from sapwood to heartwood might be related to the K content ratio of heartwood and sapwood.