Norsalsolinol uptake into secretory vesicles via vesicular monoamine transporter and its secretion by membrane depolarization or purinoceptor stimulation in PC12 cells.

The intracellular dynamics of norsalsolinol, a neurotoxin candidate causing parkinsonism-like symptoms, in PC12 cells was studied. We found that dopamine and norsalsolinol are co-localized to secretory granule layer by sucrose density gradient in norsalsolinol-treated PC12 cells. The norsalsolinol was actively taken up into isolated secretory vesicle fraction from PC12 cells with a Km value of 41.5+/-6.8 microM. The uptake of 10 microM of norsalsolinol was sensitive to reserpine (1 microM), an inhibitor of vesicular dopamine transporter, and dopamine, an endogenous substrate, but insensitive to GBR-12909, an inhibitor of dopamine transporter on plasma membrane. In norsalsolinol-treated PC12 cells, exposure to high K+ or ATP resulted in simultaneous release of norsalsolinol and dopamine. Time course of a release of dopamine and that of norsalsolinol evoked by 50 mM KCl or 100 microM ATP corresponded to each other. These releases were dependent on the concentrations of secretagogues. These data suggest that norsalsolinol is taken up with dopamine into secretory vesicle via vesicular catecholamine transporter.     

严重水分失衡状态下四照花蒸腾表面的削减

应用RGB图像分析的方法,研究了日本山口市街区生长的四照花（Comus kousa Buerg.）对2007年夏季干热多风的环境的响应。结果表明,许多四照花叶尖叶缘出现焦枯,清晰的暗棕色防护带显示在焦枯的叶片上。这些防护带逐渐从叶尖向叶基退缩直到成功控制焦枯为止,且留下许多不成功的防卫痕迹。焦枯的叶片呈现鲜明的逻辑斯蒂函数式变化趋势。依据气象数据分析,四照花叶片退缩型焦枯几乎与干燥峰值期同步。这表明在干燥度突然升高期间,极端的水分亏缺诱发了四照花的防卫反应,从而保护其余部分免于进一步的失水。像素分析结果表明,截止到2007年8月底四照花样树通过局部叶片退缩型焦枯减少了40.2%的叶面积。相比之下,鉴于上半年雨量充足,2008年,相同的样树只有13.2%叶面积减少。不管怎样,四照花的确削减了蒸腾表面积,并且表现出一种独特的蒸腾表面减少方式。既不同于落叶也不同于地上部枯死。依据脱水过程的分析,四照花在突然的干热胁迫下叶尖始发的干缩和叶片防卫反应的共同作用被认为是其蒸腾表面削减的关键所在。     

Performance of a closed recirculating system with foam separation, nitrification and denitrification units for intensive culture of eel: towards zero emission

The development of a closed recirculating aquaculture system that does not discharge effluents would reduce a large amount of pollutant load on aquatic bodies. In this study, eel were reared in a closed recirculating system, which consisted of a rearing tank, a foam separation unit, a nitrification unit and a denitrification unit. The foam separation unit has an inhalation-type aerator and supplies air bubbles to the rearing water. The growth of eel, which were fed a commercial diet, was satisfactory, with gross weight increases of up three times in 3 months. The survival rate under the congested experimental conditions was 91%. The foam separation unit maintained oxygen saturation in the rearing water at about 80%. Furthermore, fine colloidal substances were absorbed on the stable foam formed from eel mucus and were removed from the rearing water by foam separation. Ammonia oxidation and the removal of suspended solids were accomplished rapidly and simultaneously in the nitrification unit. The ammonia concentration and turbidity were kept at less than 1.2 mg of N per litre and 2.5 units, respectively. When the denitrification process was operated, nitrate that accumulated in the rearing water (151 mg of N per litre) was reduced to 40 mg of N per litre. The sludge was easily recovered from the nitrification and denitrification tanks, and the components were found suitable as compost. Based on these results, the intensive aquaculture of freshwater fish such as eel can be achieved using a closed recirculating system without emission.     

Expression of HSP70 in response to heat-shock and its cDNA cloning from Mediterranean blue mussel

Expression of HSP70 in response to heat-shock was investigated at the protein and mRNA levels in Mediterranean blue mussel. Western and Northern blot analyses revealed that HSP70 was expressed following heat-shock in the mantle at both protein and mRNA levels, suggesting that gene expression of HSP70 is implicated in the cellular response to heat-shock stress in mussel. It was then attempted to clone HSP70 cDNA in order to determine the primary structure of mussel HSP70. As a result, two full-length cDNA encoding HSP70 were isolated from a cDNA library prepared from the heat-shocked mantle. The isolated cDNA consist of single open reading frames of 2067 bp and 1911 bp which encode proteins of 689 amino acids and 637 amino acids, respectively. Both HSP70 cDNA encode an ATPase do main, and a substrate-binding do main in addition to a Glu-Glu-Val-Asp (EEVD) peptide motif that is specific for cytosolic HSP70. These findings suggest that the cDNA clones obtained in the present study encode cytosolic HSP70.     

Prevalence of Bartonella henselae, Bartonella clarridgeiae and the 16S rRNA gene types of Bartonella henselae among pet cats in Japan

The authors investigated bacteriologically the prevalence of Bartonella infection among 690 pet cats derived from 10 private animal hospitals in six cities (Sapporo, Hokkaido Prefecture; Sendai, Miyagi Prefecture; Joetsu, Niigata Prefecture; Fujisawa, Kanagawa Prefecutre; Kyoto, Kyoto Prefecture; Sanda, Hyogo Prefecutre) and 4 counties (Mishima, Osaka Prefecture; Hikawa, Shimane Prefecture; Aira, Kagoshima Prefecture; Shimajiri, Okinawa Prefecture) located from the north to the south of Japan. Bartonella species were isolated from 7.2% (50/690) of all the cats examined. No Bartonella species were isolated from the cats in Sapporo or Sendai. The isolation rate varied from 2% in Joetsu and Sanda to 20% in Shimajiri. Bartonella clarridgeiae was isolated from two of 50 cats in Kyoto, three of 50 in Mishima and one of 50 in Shimajiri, but not in cats from the other cities or counties. Though the cats of Joetsu, Fujisawa, Kyoto, Sanda, Aira and Shimajiri were infected with either B. henselae or B. clarridgeiae, one of eight infected cats in Mishima was harboring both Bartonella species. Type I of 16S rRNA gene was the predominant type among the isolates of B. henselae, but only one isolate derived from Shimajiri was found to be of type II. Prevalence of B. clarridgeiae and the 16S rRNA gene type of B. henselae among cats in Japan was demonstrated for the first time in this investigation.     

Extractives relating to heartwood color changes in sugi (<Emphasis Type="Italic">Cryptomeria japonica</Emphasis>) by a combination of smoke-heating and UV radiation exposure

 Sugi green logs with red or black heartwood were smoke-heated, and the changes in the color of the heartwood after ultraviolet (UV) (λ = 365 nm) radiation exposure were then observed. After UV radiation exposure, the redness and yellowness increased in both the red and black heartwoods, whereas the brightness decreased. In the black heartwood, the resulting color turned from yellowish white to reddish brown. Reddening in black heartwood after exposure to a combination of smoke heating and UV radiation is thought to be due to a decrease in brightness and an increase in both redness and yellowness. However, the degree of change in heartwood color by UV radiation exposure was not greatly affected by smoke-heating treatments of various lengths. When methanol extracts were fractionated and exposed to UV radiation, the yellowness increased in the n-hexane-soluble portion and the redness increased in the acetone-soluble fractions from the n-hexane-insoluble portion. These results suggest that the n-hexane-soluble fraction contains the substances that allow heartwood color to change to yellow after UV radiation exposure, and the acetone-soluble-fraction from the n-hexane-insoluble portion contains the substances that allow it to change to red. Received: November 14, 2001 / Accepted: June 3, 2002 Acknowledgment This research was supported in part by a Grant-in-Aid for Scientific Research from the Japan Society for the Promotion of Science. This study was presented in part at the 51st Annual Meeting of the Japan Wood Research Society, Tokyo, April 2001 Correspondence to:N. Yoshizawa     

Silicic acid: boric acid complexes as wood preservatives

Wood was treated with a low molecular-weight silicic acid: boric acid agent and examined for increased resistance to termites and combustion. Wood treated with silicic acid only exhibited increased termite resistance, but not to the marked extent observed after treatment with a mixture of silicic and boric acids. Increasing the quantity of boric acid also increased the termite mortality rate and shortened the time to death. Oral-toxicity tests using dyed silicic acid: boric acid suggested that the boric acid in the agent acted as a stomach poison. Field tests on stakes over three years showed that even if stakes were placed near the termite exit, those treated with silicic acid and high levels of boric acid had no attack by termites and maintained their original form. Though these specimens were installed in the field during three rainy seasons (about six months in total) in three years, the termite resistance ability of these chemicals was maintained. It became clear that this silicic acid: boric acid agent has a high water resistance ability worthy of application in the outdoors. Combustion tests showed that flame and glowing combustion times were shortened at high levels of boric acid. When a boric acid: methanol solution was added at of rate of not less than 25 ml for 100 g of colloidal silicic acid solution (CSAS), flaming and glowing combustion were not observed. Although the charring length of the wood specimen which was treated with a low molecular weight silicic acid: boric acid agent decreased to 2/3 of that of the control wood, the charring lengths were not influenced by the level of boric acid. However, the volume of smoke decreased relative to the amount of boric acid that had been added.