Emerging X-ray free-electron lasers with femtosecond pulse duration enable single-shot snapshot imaging almost free from sample damage by outrunning major radiation damage processes. In bioimaging, it is essential to keep the sample close to its natural state. Conventional high-resolution imaging, however, suffers from severe radiation damage that hinders live cell imaging. Here we present a method for capturing snapshots of live cells kept in a micro-liquid enclosure array by X-ray laser diffraction. We place living Microbacterium lacticum cells in an enclosure array and successively expose each enclosure to a single X-ray laser pulse from the SPring-8 Angstrom Compact Free-Electron Laser. The enclosure itself works as a guard slit and allows us to record a coherent diffraction pattern from a weakly-scattering submicrometre-sized cell with a clear fringe extending up to a 28-nm full-period resolution. The reconstructed image reveals living whole-cell structures without any staining, which helps advance understanding of intracellular phenomena.
Unusual long polyamines such as caldopentamine and caldohexamine, and branched polyamines such as tetrakis(3-aminopropyl)ammonium and N (4)-aminopropylspermidine were often found in cells of extreme thermophiles and hyperthermophiles belonging to both Bacteria and Archaea domains. Some of these unusual polyamines are essential for life at extreme temperatures. In some cases, the unusual polyamines also exist in cells of nonthermophilic organisms and play important physiological roles under normal conditions. Methods for chromatographic analysis, isolation, and chemical syntheses of unusual polyamines as well as experimental methods for measuring their physiological roles are discussed. Especially, many newly improved methods for chemical syntheses are presented in this article.
Two novel thermophilic micro-organisms, designated YMO81(T) and YMO722(T), were isolated from a high-temperature compost (internal temperature > 95 °C). The isolates were able to grow at 80 °C in a nutrient broth and in a synthetic medium. Cells were aerobic, Gram-negative rods (0.3 x 4.0 μm). Spore formation was not observed. Strain YMO81(T) grew at 83 °C and pH 6.9-8.9 and grew optimally at 78 °C and pH 7.5 with 2 % NaCl. For growth in a synthetic minimal medium at 70 °C, the vitamins biotin, folic acid and thiamine and the amino acids glutamine and methionine were essential for growth of both strains; at 80 °C, strain YMO81(T) also required histidine, isoleucine, leucine, lysine, phenylalanine, serine, tryptophan and valine. Cellular fatty acids of the isolates comprised mainly iso-C₁₇:₀ and anteiso-C₁₇:₀. The DNA G+C contents of strains YMO81(T) and YMO722(T) were 70 and 64 mol%, respectively. When the 16S rRNA gene sequences of the isolates were compared with those of other bacteria, highest similarity was observed with Planifilum yunnanense LA5(T) (90 % 16S rRNA gene sequence similarity). DNA-DNA relatedness between strain YMO722(T) and strain YMO81(T) was 55 %. N⁴-Aminopropylspermine was identified as a major polyamine, which suggested that the isolates were distinct from other related taxa. On the basis of phylogenetic, phenotypic and chemotaxonomic analyses, we propose a new genus, Calditerricola gen. nov., and two novel species, the type species Calditerricola satsumensis sp. nov., with type strain YMO81(T) (=ATCC BAA-1462(T) =JCM 14719(T) =DSM 45223(T)), and Calditerricola yamamurae sp. nov., with type strain YMO722(T) (=ATCC BAA-1461(T) =JCM 14720(T) =DSM 45224(T)).
We investigated the microbial community of a high-temperature compost process exhibiting an internal temperature exceeding 90 degrees C. The waste pile was crosscut and samples were collected from the bottom to the top of the refuse pile. PCR-denaturing gradient gel electrophoresis analysis suggested that the microbial community of the high-temperature compost is heterogeneous and differs from one locality to another. Heat-stable collagenases and amylases were extracted directly from the compost pile. Collagenases were located in the upper half of the pile, whereas amylases were detected mainly in the lower parts. Several extremely thermophilic strains were isolated at 80 degrees C; these strains were aerobes. Based on 16S rRNA sequence analysis, the isolates clustered together and represent one or two closely related species. We propose that these thermophilic isolates belong to a novel genus, Caldaterra, gen. nov.
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