Molecular characterization of microbial communities in fault-bordered aquifers in the Miocene formation of northernmost Japan

We investigated the diversity and distribution of archaeal and bacterial 16S rRNA gene sequences in deep aquifers of mid‐ to late Miocene hard shale located in the northernmost region of the Japanese archipelago. A major fault in the north‐west–south‐east (NW–SE) direction runs across the studied area. We collected three groundwater samples from boreholes on the south‐west (SW) side of the fault at depths of 296, 374 and 625 m below ground level (m.b.g.l.) and one sample from the north‐east (NE) side of the fault at a depth of 458 m.b.g.l. The groundwater samples were observed to be neutral and weakly saline. The total microbial counts after staining with acridine orange were in the order 10⁵?10⁶ cells mL^?1 and 10³ cells mL^?1 in the aquifers to the SW and to the NE of the fault, respectively. A total of 407 archaeal and bacterial 16S rRNA gene sequences (204 and 203 sequences, respectively) were determined for clone libraries constructed from all groundwater samples. Phylogenetic analyses showed that the libraries constructed from the SW aquifers were generally coherent but considerably different from those constructed from the NE aquifer. All of the archaeal clone libraries from the SW aquifers were predominated by a single sequence closely related to the archaeon Methanoculleus chikugoensis, and the corresponding bacterial libraries were mostly predominated by the sequences related to Bacteroidetes, Firmicutes and δ‐Proteobacteria. In contrast, the libraries from the NE aquifer were dominated by uncultured environmental archaeal clones with no methanogen sequences and by β‐proteobacterial clones with no sequences related to Bacteroidetes and δ‐Proteobacteria. Hence, the possible coexistence of methanogens and sulphate reducers in Horonobe deep borehole (HDB) on the SW side is suggested, particularly in HDB‐6 (374 m.b.g.l.). Moreover, these organisms might play an important geochemical role in the groundwater obtained from the aquifers.     

EFFECTS OF DIAMINOPROPIONATE, DEOXYADENOSINE, AND THEOPHYLLINE ON STIMULATED FORMATION OF CYCLIC AMP AND GMP BY DEPOLARIZING AGENTS IN SLICES OF GUINEA-PIG CEREBRAL CORTEX

In incubated slices of guinea-pig cerebral cortex depolarizing agents such as veratridine and high potassium ions caused 50 to 80-fold increases of adenosine 3', 5'-cyclic monophosphate (cyclic AMP) levels and these responses were inhibited about 50% by 2, 3-diaminopropionate and 2'-deoxyadenosine: the former is a specific antagonist for glutamate-elicited accumulation of cyclic AMP and the latter selectively for adenosine-elicited accumulation. Methylxanthines were powerful ‘inhibitors’toward the responses not only to depolarizing agents but also to glutamate and adenosine. These findings are consistent with the hypothesis that releases of both glutamate and adenosine are involved in the depolarization-elicited increases of cyclic AMP levels. Guanosine 3', 5'-cyclic monophosphate (cyclic GMP) levels in the slices were also elevated by veratridine as well as by glutamate, but always to a lesser extent (8 ～ 12 times the control value) than cyclic AMP levels were. The responses for cyclic GMP both to veratridine and glutamate were ‘augmented’by methylxanthines and were not inhibited by 2, 3-diaminopropionate. Thus, glutamate appears to cause the increase of cyclic GMP levels through a different mechanism or site of action from that for cyclic AMP.     

Distinctive developmental variability of genital parts in the sexually dimorphic beetle, Prosopocoilus inclinatus (Coleoptera: Lucanidae)

Recent comparative studies have revealed that the rapid diversity of genitalia is closely related to sexual selection and that genital development interacts with the development of different body parts. Hypotheses about developmental stability due to selection to genital parts were tested by estimating allometric relations in a sexually dimorphic stag beetle Prosopocoilus inclinatus . All genital parts of males scaled to body size with a slope of less than 1 and all but the median lobe (male intromittent organ) showed smaller variability than other body parts. This supported the 'one-size-fits-all' hypothesis, which suggests broad copulation opportunity by males of any size with females within a population. Nevertheless, we found large variation among different genital parts in coefficients of variation and in values of the switch point where the allometric relations varied significantly. These results strongly support the view that developmental trajectories of genital traits are not necessarily integrated. Among the genitalic traits, male intromittent organ and female genitalia exhibited large variability, suggesting a high responsiveness to the selective regimes and physical interaction during copulation. This may account for rapid diversification of genital morphology, even in closely-related populations in beetle species.  © 2007 The Linnean Society of London, Biological Journal of the Linnean Society , 2007, 90 , 573–581.     

Action Spectra for Step-Up Photophobic Response in Blepharisma

ABSTRACT The cells of Blepharisma which possess red pigment (blepharismin) show step-up photophobic response (temporal ciliary reversal induced by a sudden increase in light intensity). Bleaching of the cells by cold shock raised a threshold light intensity for the response, Oxidation of red pigment that produced blue pigment did not raise the threshold for the response. The action spectrum for the step-up photophobic response of the cells which possess normal red pigment had peaks at about 580, 540 and 490 nm, a value which coincided with peaks of an absorption spectrum of the red pigment. The absorption spectrum of oxidized pigment (blue pigment) shifted 20 nm toward infrared light. The action spectrum for the response of the cells which possess blue pigment also shifted 20 nm toward infrared light. Results suggest that red pigment might be involved in the step-up photophobic response. Key words. Blepharismin, ciliary reversal, photoreceptors, photoresponse.     

Effects of lifestyle and genetic polymorphisms on consumption of coffee or black tea and urinary caffeine levels

To find the cause of individual differences in caffeine intake and its metabolism, we investigated the effects of lifestyle and genetic polymorphisms of caffeine metabolic enzymes on coffee or black tea and urinary caffeine levels among 259 male Japanese. It was seen that cigarette smokers drank more coffee or black tea than non-smokers (p < 0.001). There was an inverse correlation between the amount of coffee or black tea consumed and age or the frequency of alcohol drinking (p < 0.05). Genetic polymorphisms of N -acetyltransferase 2 (NAT2), cytochrome P450 (CYP)1A1 and 2E1 did not significantly affect the habit of drinking coffee or black tea. The frequency of allele 1, the NAT2 allele of rapid acetylators, increased according to coffee or black tea consumption (0.05 < p < 0.1). Among lifestyle factors, two factors, i.e. smoking and the amount of coffee or black tea consumed, were related to urinary caffeine levels (p < 0.05). Geometric means of urinary caffeine levels were higher in the group who consumed higher amounts of coffee or black tea (p < 0.05) and those of smokers were lower than non-smokers- approximately 70% of non-smokers (p < 0.05). The genetic polymorphisms of NAT2, CYP1A1 and CYP2E1 were not significantly associated with the urinary caffeine levels according to each consumption level of coffee or black tea. This study suggests that smoking should be considered for the proper appreciation of individual differences in caffeine intake and urinary caffeine levels.     

Temperature-Sensitive Responses in Blepharisma

Temperature sensitivity of Blepharisma cultured at 23°C was investigated in a temperature range between 18.5°C and 33.5°C. The cells accumulated in an optimal temperature (ca. 27°C) region when they were placed in a chamber with a temperature gradient, although a certain population of the cells accumulated at much higher temperatures. The quantitative analysis of behavioral responses exhibited by the cells revealed that three types of thermal response were responsible for thermoaccumulation of the cells in an optimal temperature: (1) an increase in the frequency of thermophobic response in the cells swimming away from the optimal temperature region; (2) acceleration of forward swimming velocity of the cells swimming toward the optimal temperature region; and (3) higher frequency of spontaneous ciliary reversal of the cells in higher temperature regions.     

ONTOGENY OF CHICKEN HEMOGLOBIN. III. IMMUNOLOGICAL STUDY OF THE HETEROGENEITY OF HEMOGLOBIN IN DEVELOPMENT

By applying the double diffusion technique of Ouchterlony and the immunoelectrophoresis, sequence in the appearance of antigens reactive with antisera against HbCOs from 5-day embryos and adult chickens, and the major component of adult HbCO in the course of chicken development has been studied.
Antigenic components reacting to antiserum against HbCO from 5-day embryos have been detected throughout development. Three globin-like components specific to early embryos are detectable in embryos to 2 days of incubation. Two Hb components are detectable in embryos from 3 to 5 days of incubation; one is apparently specific to embryos, while the other seems to be somewhat different from adult Hb. After 6 days of incubation only one adult Hb component is detectable.
Antigenic components reacting to antiserum against HbCO from adults have also been detectable throughout embryonic life. One globin-like component specific to early embryos can be found in embryos to 2 days of incubation. One or two Hb components which are probably specific to embryos can be detected in embryos from 3 to 5 days of incubation. After 6 days of incubation one adult Hb component is detectable, while one globin-like component specific to adults can be found after 15 days of incubation. Further, the other globin-like component is detectable after 3 days of incubation.
Antigenic components reacting to antiserum against the major component of adult HbCO have been detected throughout development; one which seems to be somewhat different from either adult Hb components can be found in embryos from 2 to 5 days of incubation, while the other which is identical with the major component of adult Hb is detectable after 6 days of incubation.     

神经降压素对迷走运动背核神经元兴奋作用的离体研究

用细胞内微电极记录方法在大鼠离体脑片中研究了神经降压素(NT)对迷走运动背核(DMV)神经元的作用机制。滴注或灌流NT使88％DMV神经元产生剂量依赖性去极化反应,并伴有膜阻抗的增加。去极化反应的翻转电位约-82mV,反应幅度受灌流液中钾离子浓度的影响。用高镁无钙液或含河豚毒素的灌流液阻断突触传递后,去极化反应仍然存在。研究结果提示NT通过突触后机制直接兴奋DMV神经元,此去极化兴奋作用可能与钾通道的关闭有关。