Distinct kinetic changes in neurotransmitter release after SNARE protein cleavage

Neurotransmitter release is triggered by calcium ions and depends critically on the correct function of three types of SNARE [soluble N-ethylmaleimide-sensitive factor attachment protein (SNAP) receptor] proteins. With use of the large calyx of Held presynaptic terminal from rats, we found that cleavage of different SNARE proteins by clostridial neurotoxins caused distinct kinetic changes in neurotransmitter release. When elevating calcium ion concentration directly at the presynaptic terminal with the use of caged calcium, cleavage of SNAP-25 by botulinum toxin A (BoNT/A) produced a strong reduction in the calcium sensitivity for release, whereas cleavage of syntaxin using BoNT/C1 and synaptobrevin using tetanus toxin (TeNT) produced an all-or-nothing block without changing the kinetics of remaining vesicles. When stimulating release by calcium influx through channels, a difference between BoNT/C1 and TeNT emerged, which suggests that cleavage of synaptobrevin modifies the coupling between channels and release-competent vesicles.     

Mass spectrometry of intact V-type ATPases reveals bound lipids and the effects of nucleotide binding

The ability of electrospray to propel large viruses into a mass spectrometer is established and is rationalized by analogy to the atmospheric transmission of the common cold. Much less clear is the fate of membrane-embedded molecular machines in the gas phase. Here we show that rotary adenosine triphosphatases (ATPases)/synthases from Thermus thermophilus and Enterococcus hirae can be maintained intact with membrane and soluble subunit interactions preserved in vacuum. Mass spectra reveal subunit stoichiometries and the identity of tightly bound lipids within the membrane rotors. Moreover, subcomplexes formed in solution and gas phases reveal the regulatory effects of nucleotide binding on both ATP hydrolysis and proton translocation. Consequently, we can link specific lipid and nucleotide binding with distinct regulatory roles.     

Structure of the rotor of the V-Type Na+-ATPase from Enterococcus hirae

The membrane rotor ring from the vacuolar-type (V-type) sodium ion-pumping adenosine triphosphatase (Na+-ATPase) from Enterococcus hirae consists of 10 NtpK subunits, which are homologs of the 16-kilodalton and 8-kilodalton proteolipids found in other V-ATPases and in F1Fo- or F-ATPases, respectively. Each NtpK subunit has four transmembrane alpha helices, with a sodium ion bound between helices 2 and 4 at a site buried deeply in the membrane that includes the essential residue glutamate-139. This site is probably connected to the membrane surface by two half-channels in subunit NtpI, against which the ring rotates. Symmetry mismatch between the rotor and catalytic domains appears to be an intrinsic feature of both V- and F-ATPases.     

Quantitative nested real-time PCR detection of <Emphasis Type="Italic">Verticillium longisporum</Emphasis> and <Emphasis Type="Italic">V. dahliae</Emphasis> in the soil of cabbage fields

Verticillium longisporum and V. dahliae, causal agents of Verticillium wilt, are spreading through the cabbage fields of Gunma Prefecture. Using the V. longisporum-specific intron within the 18S rDNA and differences between ITS 5.8S rDNA sequences in Japanese isolates of V. longisporum and V. dahliae, we developed three quantitative nested real-time (QNRT) PCR assays. The QNRT-PCR quantification of V. longisporum or V. dahliae in cabbage field soil was consistent with the severity of Verticillium wilt disease in those fields. In field trials of resistant cultivar YR Ranpo grown for three seasons in soil infested with the pathogen, disease severity and pathogen density in the soil were significantly reduced in a field moderately contaminated by V. dahliae, but only slightly reduced in a highly contaminated field. These results suggest that continuous cultivation of a resistant cultivar is an effective way to reduce the pathogen population. QNRT-PCR assays provide a powerful analytical tool to evaluate the soil population dynamics of V. longisporum and V. dahliae for disease management.     

评价大米储藏品质的炊饭特性指标的研究

提出了评价大米储藏品质的炊饭特性新指标——光透过率差。实验证明，随着大米品质的劣化，光透过率差明显变小。以此指标为主，结合原有的加热吸水率和ｐＨ值，对因生霉和陈化引起的大米储藏品质的变化，都能较好地进行评价。同时对影响这些指标稳定的实验因素也进行了探讨。     

Effect of rapid oligotrophication by an aquatic treatment pilot plant on the microbial community of a mesotrophic Bog

Rapid oligotrophication by the operation of the ‘Bio-filter system’ has changed Doh-Hoh-Numa Bog from a moderate mesotrophic system to the oligotrophic end of such a system. However, the population growth rate of the bacterioplankton community was little affected, and continued to be controlled primarily by the DOC concentration even after the system operation. This DOC concentration was higher during the operation period, although environmental factors relevant to primary production were shifted to the direction of oligotrophication. The natural bacterioplankton community in the bog was shown to modify its physiological kinetics to maintain the growth rate at the optimal level, even when it was exposed to oligotrophication processes with the operation of the ‘Bio-filter system’.     

Food-drug interaction between ferulic acid and nateglinide involving the fluorescein/H+ cotransport system

In clinical, patients usually take many kinds of drugs at the same time. Thus, drug-drug interactions involving transporters can often directly affect the therapeutic safety and efficacy of many drugs. However, there have been few studies on food-drug interactions involving transporters. Dietary polyphenols have been widely assumed to be beneficial to human health. Polyphenols are commercially prepared and used as functional foods. We report here for the first time that ferulic acid, which is widely used as a functional food, affects the transport of clinical agents. It is important to be aware of the potential of food-drug interactions and to act in order to prevent undesirable and harmful clinical consequences.     

Growth, Net Photosynthetic Rate, Nutrient Status and Secondary Xylem Anatomical Characteristics of Fagus Crenata Seedlings Grown in Brown Forest Soil Acidified with H2SO4 Solution

Dry matter production, net photosynthetic rate, leaf nutrient status and trunk anatomical characteristics of Fagus crenata seedlings grown in brown forest soil acidified by adding H₂SO₄ solution were investigated. The soil acidification leaded to decreased (Ca+Mg+K)/Al molar ratio in the soil solution. Dry mass per plant of the seedlings grown in the soil treated with H⁺ at 120 mg·L^?1 was significantly reduced compared with the control value at 0 mg·L^?1. When net photosynthetic rate was reduced in the seedlings grown in the soil treated with H⁺ at 120 mg·L^?1, the carboxylation efficiency and maximum net photosynthetic rate at saturated CO₂-concentration were lower than the control values. The addition of H⁺ to the soil at 120 mg·L^?1iinduced a reduction in the concentration of Ca in the leaf. By contrast, the concentration of Al in the leaf was increased with increasing the amount of H⁺ added to the soil. The annual ring formed in the seedlings grown in the soil treated with H⁺ at 120 mg·L^?1 was significantly narrower than that at 0 (control), 10, 30, 60 or 90 mg·L^?1. Based on the results obtained in the present study, we conclude that Fagus crenata is relatively sensitive to a reduction in the (Ca+Mg+K)/Al molar ratio of soil solution compared with Picea abies.     

Supercritical carbon dioxide extraction of turmeric (Curcuma longa)

Turmeric oil was extracted from turmeric (Curcuma longa) with supercritical carbon dioxide in a semicontinuous-flow extractor. Extraction rate was measured as a function of pressure, temperature, flow rate, and particle size. The extraction rate increased with an increase in CO(2) flow rate and with a reduction of particle size. The effect of pressure and temperature on turmeric extraction suggested the use of higher pressure and lower temperature at which solvent density is greater and thus the solubility of the oil in the solvent is greater in the range of 313-333 K and 20-40 MPa. The major components ( approximately 60%) of the extracted oil were identified as turmerone and ar-turmerone by GC-MS.