微生物マイクロハビタットとしての土壌団粒微小環境の解明

科学研究費助成事業 研究成果報告書：若手研究(B)2015-2017課題番号 : 15K1865

Contactless mass transfer for intra-droplet extraction

This study demonstrates the possibility of “contactless” mass transfer between two aqueous slugs (droplets) separated by an oil slug in Taylor flow inside milli-channels. Separation of the alternating aqueous slugs at the outlet was performed by switching a couple of solenoid valves at branched outlets according to signals obtained by an optical sensor at the branch. Transfer of bromothymol blue (BTB) from acidic to basic aqueous slugs was performed for demonstration. In some cases, aqueous slugs separated by oil, merged catching on each other due to the velocity difference. Interfacial tension which was affected by the solute concentration was responsible for the velocity difference. Position-specific mass transfer activity at the rear end of the aqueous slugs was found on the course of the experiment. A meandering channel decreased the velocity difference and enhanced mass transfer. Almost complete (93%) transfer of BTB was achieved within a short residence time of several minutes under optimized conditions. The presented system opens a way for advanced separation using minimum amounts of the oil phase and allows concentrating the solute by altering relative lengths of the sender and receiver slugs

Distinctive Roles of Two Aggregate Binding Agents in Allophanic Andisols: Young Carbon and Poorly-Crystalline Metal Phases with Old Carbon

Interaction of organic matter (OM) with soil mineral components plays a critical role in biophysical organization (aggregate structure) as well as in biogeochemical cycling of major elements. Of the mineral components, poorly-crystalline phases rich in iron (Fe) and aluminum (Al) are highly reactive and thus contribute to both OM stabilization and aggregation. However, the functional relationship among the reactive metal phases, C stability, and aggregation remains elusive. We hypothesized that relatively young C acts as a binding agent to form the aggregates of weak physical stability, whereas the reactive metal phases and older C bound to them contribute to stronger aggregation. Using four surface horizons of Andisols having a gradient of soil C concentration due to decadal OM management, we conducted sequential density fractionation to isolate six fractions (from 2.5 g cm−3) with mechanical shaking, followed by selective dissolution and radiocarbon analysis. After 28 years of no-till with litter compost addition, not only C and N but inorganic materials including the reactive metal phases (pyrophosphate-, oxalate-, and dithionite-extractable metals) showed clear shifts in their concentrations towards lower-density fractions (especially <2.0 g cm−3) on a ground-area basis. This result was explained by the binding of compost-derived OM with soil particles. Major portions of the reactive metal phases in bulk samples were distributed in mid-density fractions (2.0–2.5 g cm−3) largely as sonication-resistant aggregates. Theoretical density calculations, together with depletion in radiocarbon (Δ14C: −82 to −170‰) and lower C:N ratio, implied that the sorptive capacity of the reactive metal phases in these fractions were roughly saturated with pre-existing OM. However, the influx of the compost-derived, modern C into the mid-density fractions detected by the paired-plot comparison suggests decadal C sink in association with the reactive metal phase. Our results supported the concept of aggregate hierarchy and further provided the following new insights. At the high hierarchy level where shaking-resistant aggregates form, soil organo-mineral particles appeared to be under a dynamic equilibrium and the changes in OM input regime controlled (dis)aggregation behavior due to the binding effect of relatively young C. At a lower hierarchy level, the reactive metal phases were bound to N-rich, 14C-depleted OM and together functioned as persistent binding agent. Our study suggests that the recognition of binding agents and aggregate hierarchy level would help to untangle the complex organo-mineral interactions and to better understand soil C stability

The Nutritional Status of Japanese Black Breeding Cows Grazed in a Larger Abandoned Paddy Field

Because rice consumption has decreased in Japan, the number of abandoned paddy fields has increased. Some of the abandoned paddy fields are being used for livestock grazing, effectively using the abandoned fields and reducing the production costs on livestock farms. Such grazing may use an integrated grazing system of scattered small pastures where each of the small abandoned paddy fields scattered in a given agriculture area is defined as one grazing yard, and a herd consisting of two or three head of cattle is moved from one grazing yard to another. However, because the number of abandoned paddy fields has increased, it may be possible to have cattle grazing in larger fields made by combining multiple abandoned paddy fields (Shindo and Tejima 2006). Here we examined the nutritional status of Japanese Black breeding cows that stayed in larger-sized abandoned paddy fields to graze from spring to autumn

Silenced Expression of NFKBIA in Lung Adenocarcinoma Patients with a Never-smoking History

Nuclear factor of κ-light polypeptide gene enhancer in B cells inhibitor α (NFKBIA), which is a tumor suppressor gene, was found to be silenced in lung adenocarcinomas. We examined NFKBIA expression, mutations in the EGFR and K-ras genes, and EML4-ALK fusion in 101 resected lung adenocarcinoma samples from never-smokers. NFKBIA expression was evaluated using immunohistochemistry. NFKBIA expression was negative in 16 of the 101 samples (15.8%). EGFR and K-ras mutations and EML4-ALK fusion were detected in 61 (60.5%), 1 (1.0%), and 2 (2.0%) of the 101 samples, respectively, in a completely mutually exclusive manner. Negative NFKBIA expression was observed significantly more frequently among the tumors with none of the three genetic alterations compared to those with such alterations (p＝0.009). In addition, negative NFKBIA expression was significantly more frequent among the EGFR-wild type samples compared to the EGFR-mutant samples (p＝0.013). In conclusion, NFKBIA expression was silenced in adenocarcinomas without EGFR/K-ras mutations or EML4-ALK fusion, suggesting that the silencing of NFKBIA may play an important role in the carcinogenesis of adenocarcinomas independent of EGFR/K-ras mutations or EML4-ALK fusion

Musical instrument training program improves verbal memory and neural efficiency in novice older adults

楽器訓練で高齢者の認知機能が向上することを確認 --訓練による脳活動の変化を高齢者で初報告--. 京都大学プレスリリース. 2020-12-24.Previous studies indicate that musical instrument training may improve the cognitive function of older adults. However, little is known about the neural origins of training‐related improvement in cognitive function. Here, we assessed the effects of instrumental training program on cognitive functions and neural efficiency in musically naïve older adults (61–85 years old). Participants were assigned to either the intervention group, which received a 4‐month instrumental training program using keyboard harmonica, or a control group without any alternative training. Cognitive measurements and functional magnetic resonance imaging during visual working memory (VWM) task were administered before and after the intervention in both groups. Behavioral data revealed that the intervention group significantly improved memory performance on the test that measures verbal recall compared to the control group. Neuroimaging data revealed that brain activation in the right supplementary motor area, left precuneus, and bilateral posterior cingulate gyrus (PCgG) during the VWM task decreased after instrumental training only in the intervention group. Task‐related functional connectivity (FC) analysis revealed that the intervention group showed decreased FC between the right PCgG and left middle temporal gyrus, and between the left putamen and right superior temporal gyrus (lPu‐rSTG) during a VWM task after the intervention. Furthermore, a greater improvement in memory performance in the intervention group was associated with a larger reduction in lPu‐rSTG FC, which might be interpreted as improved neural efficiency. Our results indicate that the musical instrument training program may contribute to improvements in verbal memory and neural efficiency in novice older adults

In Search of a Binding Agent: Nano-Scale Evidence of Preferential Carbon Associations with Poorly-Crystalline Mineral Phases in Physically-Stable, Clay-Sized Aggregates

Mechanisms of protecting soil carbon (C) are still poorly understood despite growing needs to predict and manage the changes in soil C or organic matter (OM) under anticipated climate change. A fundamental question is how the submicron-scale interaction between OM and soil minerals, especially poorly-crystalline phases, affects soil physical aggregation and C stabilization. Nano-sized composites rich in OM and poorly-crystalline mineral phases were presumed to account for high aggregate stability in the Andisol we previously studied. Here we searched for these nanocomposites within a sonication-resistant aggregate using scanning transmission X-ray microscopy (STXM) and near-edge X-ray absorption fine structure (NEXAFS) as well as electron microscopy (SEM, TEM). Specifically, we hypothesized that nanometer-scale spatial distribution of OM is controlled by poorly-crystalline minerals as both co-exist as physically-stable nanocomposites. After maximum dispersion of the cultivated Andisol A-horizon sample in water, one aggregate (a few p.m in diameter) was isolated from 0.2-2 mu m size fraction which accounted for 44-47% of total C and N and 50% of poorly-crystalline minerals in bulk soil. This fraction as well as 2 mu m size fractions, implying high abundance of the nanocomposites in the smaller fractions. The isolated aggregate showed a mosaic of two distinctive regions. Smooth surface regions showed low adsorption intensity of carbon K-edge photon energy (284-290 eV) with well-crystalline mineralogy, whereas rough surface regions had features indicative of the nanocomposites: aggregated nanostructure, high C intensity, X-ray amorphous mineral phase, and the dominance of Si, O, Al, and Fe based on SEM/EDX and TEM/EDX. Carbon functional group chemistry assessed by NEXAFS showed the dominance of amide and carboxyl C over aromatic and aliphatic C with some variation among the four rough surface regions. Together with C and N isotopic patterns among the size fractions (relatively low C:N ratio, high N-15 natural abundance, and more positive Delta C-14 of the <2 mu m fractions), our results provided the direct evidence of preferential binding of microbially-altered, potentially-labile C with poorly-crystalline mineral phases at submicron scale. The role of the nanocomposite inferred from this study may help to bridge the knowledge gap between physical aggregation process and biogeochemical reactions taking place within the soil physical structure

Gene up-regulation in response to predator kairomones in the water flea, Daphnia pulex

<p>Abstract</p> <p>Background</p> <p>Numerous cases of predator-induced polyphenisms, in which alternate phenotypes are produced in response to extrinsic stimuli, have been reported in aquatic taxa to date. The genus <it>Daphnia </it>(Branchiopoda, Cladocera) provides a model experimental system for the study of the developmental mechanisms and evolutionary processes associated with predator-induced polyphenisms. In <it>D. pulex</it>, juveniles form neckteeth in response to predatory kairomones released by <it>Chaoborus </it>larvae (Insecta, Diptera).</p> <p>Results</p> <p>Previous studies suggest that the timing of the sensitivity to kairomones in <it>D. pulex </it>can generally be divided into the embryonic and postembryonic developmental periods. We therefore examined which of the genes in the embryonic and first-instar juvenile stages exhibit different expression levels in the presence or absence of predator kairomones. Employing a candidate gene approach and identifying differentially-expressed genes revealed that the morphogenetic factors, <it>Hox3</it>, <it>extradenticle </it>and <it>escargot</it>, were up-regulated by kairomones in the postembryonic stage and may potentially be responsible for defense morph formation. In addition, the juvenile hormone pathway genes, <it>JHAMT </it>and <it>Met</it>, and the insulin signaling pathway genes, <it>InR </it>and <it>IRS-1</it>, were up-regulated in the first-instar stage. It is well known that these hormonal pathways are involved in physiological regulation following morphogenesis in many insect species. During the embryonic stage when morphotypes were determined, one of the novel genes identified by differential display was up-regulated, suggesting that this gene may be related to morphotype determination. Biological functions of the up-regulated genes are discussed in the context of defense morph formation.</p> <p>Conclusions</p> <p>It is suggested that, following the reception of kairomone signals, the identified genes are involved in a series of defensive phenotypic alterations and the production of a defensive phenotype.</p

A marine phytoplankton (Prymnesium parvum) up-regulates ABC transporters and several other proteins to acclimatize with Fe-limitation

Iron (Fe) is one of the vital limiting factors for phytoplankton in vast regions of the contemporary oceans, notably the high nutrient low chlorophyll regions. Therefore, it is apparent to be acquainted with the Fe uptake strategy of marine phytoplankton under Fe-limited condition. In the present study, marine phytoplankton Prymnesium parvum was grown under Fe-deplete (0.0025 μM) and Fe-rich (0.05 μM) conditions, and proteomic responses of the organism to Fe conditions were compared. In sodium dodecyl sulfate (SDS) gel electrophoresis, 7 proteins (16, 18, 32, 34, 75, 82, and 116. kDa) were highly expressed under Fe-deplete condition, while one protein (23. kDa) was highly expressed under Fe-rich condition. These proteins were subjected to 2-dimensional gel electrophoresis (2-D DIGE) to differentiate individual proteins, and were identified by matrix-assisted laser desorption-ionization-time of flight-mass spectrometer (MALDI-TOF-MS) analysis. The results showed that under Fe-deplete condition P. parvum increases the biosynthesis of ATP binding cassette (ABC) transporters, flagellar associated protein (FAP), and Phosphoribosylaminoimidazole-succinocarboxamide synthase. These proteins are assumed to be involved in a number of cellular biochemical processes that facilitate Fe acquisition in phytoplankton. Under Fe-deplete condition, P. parvum increases the synthesis of ribulose biphosphate carboxylase (RuBisCo), malate dehydrogenase, and two Fe-independent oxidative stress response proteins, manganese superoxide dismutase (MnSOD) and Serine threonine kinase (STK). Thus, marine phytoplankton may change their Fe acquisition strategy by altering the biosynthesis of several proteins in order to cope with Fe-limitation. © 2013 Elsevier Ltd