New monitoring approach for metabolic dynamics in microbial ecosystems using stable-isotope-labeling technologies

We have developed a new approach for monitoring the metabolic dynamics in microbial ecosystems using a combination of DNA fingerprinting and metabolome analysis based on stable-isotope-labeling technologies. Stable-isotope probing of DNA (DNA-SIP) has been used previously for the evaluation of cross-feeding in microbial communities. For the development and validation of our monitoring approach, fecal microbiota were analyzed with stable-isotope-labeled glucose used as the sole carbon source. In order to link the metabolic information and the microbial variability, we performed metabolic–microbial correlation analysis based on nuclear magnetic resonance (NMR) profiles and denaturing gradient gel electrophoresis (DGGE) fingerprints, which successfully identified the glucose-utilizing bacteria and their related extracellular metabolites. Moreover, our approach revealed information regarding the carbon flux, in that the “first” wave of extracellular metabolites secreted by the glucose-utilizing bacteria were incorporated into the “secondary” group of substrate-utilizing bacteria, and that this “secondary” group further produced their own secondary metabolized substrates. Thus, this approach is a powerful tool for monitoring the metabolic dynamics in microbial ecosystems and allows for the tracking of the carbon flux within a microbial community.     

Chiral sensing system based on the formation of diastereomeric metal complex on a homocysteine monolayer using field effect transistor

A self-assembled monolayer (SAM) of l-homocysteine (l-Hcy) formed on the surface of a gold-deposited gate of a field effect transistor (FET) was used to differentiate between enantiomers of amino acids, for which the formation of diastereomeric metal complexes is fundamental for chiral discrimination. Here, we focus our attention on the dependence of the FET response on the analyte amino acids, the central metal ions involved in complex formation, and the solution pH. Using the l-Hcy SAM-modified gate with added Cu(II), notable negative FET responses were enantioselectively observed for the l-enantiomers of alanine (Ala), phenylalanine, and tryptophan, whereas differences in the FET responses between enantiomers were negligible for asparagine and aspartic acid. Regarding the enantioselectivity for Ala, the addition of Cu(II) was demonstrated to show higher selectivity as compared to other metal ions such as Co(II) and Ni(II). Moreover, for the addition of l-Ala and Cu(II), a particularly strong negative FET response was observed at pH 5.5.     

Effect of surface roughness and surface modification of indium tin oxide electrode on its potential response to tryptophan

The effect of surface modification of indium tin oxide (ITO) electrode on its potential response to tryptophan was investigated for ITO substrates with different surface roughness. It was found that a small difference in surface roughness, between ∼1 and ∼2 nm of R_a evaluated by atomic force microscopy, affects the rest potential of ITO electrode in the electrolyte. A slight difference in In:Sn ratio at the near surface of the ITO substrates, measured by angle-resolved X-ray photoelectron spectrometry and Auger electron spectroscopy is remarkable, and considered to relate with surface roughness. Interestingly, successive modification of the ITO surface with aminopropylsilane and disuccinimidyl suberate, of which essentiality to the potential response to indole compounds we previously reported, improved the stability of the rest potential and enabled the electrodes to respond to tryptophan in case of specimens with R_a values ranging between ∼2 and ∼3 nm but not for those with R_a of ∼1 nm. It was suggested that there are optimum values of effective work function of ITO for specific potential response to tryptophan, which can be obtained by the successive modification of ITO surface.     

Amygdala beta-noradrenergic influence on hippocampal long-term potentiation in vivo

We have previously shown that hippocampal long-term potentiation (LTP), one form of synaptic plasticity that may underlie learning and memory, is attenuated by blocking neuron activity of the basolateral amygdala (BLA). In the present study we investigated the amygdala noradrenergic or cholinergic contribution to hippocampal LTP formation. When propranolol, a beta-adrenoceptor antagonist, was injected into the BLA 10 min before tetanus, the formation of LTP in the perforant path-dentate granule cell synapses was significantly impaired. Scopolamine, a muscarinic cholinergic receptor antagonist, did not affect the formation of LTP. These results suggest that amygdala beta-noradrenergic activity plays a critical role in modulation of hippocampal LTP.     

A novel antagonist, No. 7943, of the Na+/Ca2+ exchange current in guinea-pig cardiac ventricular cells

1. The effects of No. 7943 on the Na+/Ca2+ exchange current and on other membrane currents were investigated in single cardiac ventricular cells of guinea-pig with the whole-cell voltage-clamp technique. 2. No. 7943 at 0.1-10 microM suppressed the outward Na+/Ca2+ exchange current in a concentration-dependent manner. The suppression was reversible and the IC50 value was approximately 0.32 microM. 3. No. 7943 at 5-50 microM suppressed also the inward Na+/Ca2+ exchange current in a concentration-dependent manner but with a higher IC50 value of approximately 17 microM. 4. In a concentration-response curve, No. 7943 raised the K(m)Ca2+ value, but did not affect the Imax value, indicating that No. 7943 is a competitive antagonist with external Ca2+ for the outward Na+/ Ca2+ exchange current. 5. The voltage-gated Na+ current, Ca2+ current and the inward rectifier K+ current were also inhibited by No. 7943 with IC50S of approximately 14, 8 and 7 microM, respectively. 6. In contrast to No. 7943, 3', 4'-dichlorobenzamil (DCB) at 3-30 microM suppressed the inward Na+/Ca2+ exchange current with IC50 of 17 microM, but did not affect the outward exchange current at these concentrations. 7. We conclude that No. 7943 inhibits the outward Na+/Ca2+ exchange current more potently than any other currents as a competitive inhibitor with external Ca2+. This effect is in contrast to DCB which preferentially inhibits the inward rather than the outward Na+/Ca2+ exchange current.     

MR imaging of the uterus: findings from high-resolution multisection dynamic imaging with a surface coil

MR imaging of the female pelvis with a local coil (multiple phased array coils) has been reported. With this method, the signal-to-noise ratio (S/N) is improved so that high-resolution images can be obtained. Dynamic contrast-enhanced MR imaging has been reported to be useful in diagnosing uterine neoplasms. However, dynamic MR imaging is done in only limited imaging planes. When multisection dynamic MR imaging with fast gradient-echo sequences is combined with the use of a local coil, high-resolution images can be obtained throughout the entire uterus during a single breath-hold. This pictorial essay illustrates the appearance of the normal uterus and malignant uterine neoplasms on high-resolution multisection dynamic MR images obtained with a Helmholtz-type surface coil.     

Solid-State Polymerization Behavior of 1,3-Bis(3-quinolyl)-1,4-butadiyne

Summary 1,4-Bis(3-quinolyl)-1,3-butadiyne (DQ) is known to be polymerized in solid state to give the corresponding polydiacetylene. However, the polymer yield of DQ bulk crystals is low. Thus, we prepared several types of DQ crystals by different procedures to find the reason for the low polymer yield. We found three modifications of DQ crystals and they were evaluated by spectroscopic measurements and X-ray diffraction. DQ bulk crystals (Crystal I) and thermally grown DQ crystals on Crystal I (Crystal II) have the same structure classified to Type A, which gives regular polydiacetylene structure in low polymer yields. DQ crystals grown on glass plates by sublimation (Crystal III) belong to the second modification of Type B. DQ nanocrystals prepared by the reprecipitation method (Crystal IV) are Type B, and thermally grown DQ crystals on nanocrystals (Crystal V) are Type C. Crystals of Types B and C could be polymerized in low yields without showing excitonic absorption of polydiacetylene indicating irregular polymerization other than 1,4-addition. For crystals of Type A, we found that the 1,4-addition polymerization proceeded only in near-surface portions of the crystals. It can be plausibly explained that mobile monomers in the near-surface portion are only able to take part in their polymerization, resulting in low polymer conversion.     

Effect of transcutaneous electrostimulation on noise-induced temporary threshold shift

The effect of transcutaneous electrostimulation around the ear before and during noise exposure on noise-induced temporary threshold shift (TTS) was examined in 26 volunteers. Electrostimulation reduced TTS in the majority of cases and the reduction was statistically significant. Two possible mechanisms for this reduction are proposed: stimulation of the olivocochlear bundle and alteration of cochlear blood flow. Transcutaneous electrostimulation may be useful for prevention or treatment of noise induced hearing damage and for treatment of tinnitus.     

Influences of electroosmotic flows in nanopillar chips on DNA separation: Experimental results and numerical simulations

The various potential factors affecting the performance of nanopillar chips on DNA separation were investigated from the viewpoints of both numerical calculations and actual experiments. To yield higher performance and replace the conventional DNA separation techniques such as microchip electrophoresis, the phenomenon specific to the nanopillar chips should be deeply understood. In this paper, although various factors affecting the performance of the nanopillar chips are considered, we focused on the effect of electroosmotic flow, which is particularly noticeable in quartz-made nanopillar chips. High-resolution separation of DNA was realized when an electroosmotic flow was suppressed by simply using a higher concentration of buffer, but DNA separation failed in the presence of an electroosmotic flow. It was confirmed from the numerical simulations and the direct observations that the deformation of DNA band during the injection process was induced by electroosmotic flow and consequently led to a poor resolution.