ALBERT

Description: Tail-anchored (TA) proteins, a class of membrane proteins having an N-terminal cytoplasmic region anchored to the membrane by a single C-terminal transmembrane domain, are posttranslationally inserted into the endoplasmic reticulum (ER) membrane. In yeasts, the posttranslational membrane insertion is mediated by the Guided Entry of TA Proteins (GET) complex. Get3, a cytosolic ATPase, targets newly synthesized TA proteins to the ER membrane, where Get2 and Get3 constitute the Get3 receptor driving the membrane insertion. While mammalian cells employ TRC40 and WRB, mammalian homologs of Get3 and Get1, respectively, they lack the gene homologous to Get2. We recently identified calcium-modulating cyclophilin ligand (CAML) as a TRC40 receptor, indicating that CAML was equivalent to Get2 in the context of the membrane insertion. On the other hand, CAML has been well characterized as a signaling molecule that regulates various biological processes, raising the question of how the two distinct actions of CAML, the membrane insertion and the signal transduction, are assembled. In this review, we summarize recent progress of the molecular mechanism of the membrane insertion of TA proteins and discuss the possibility that CAML could sense the various signals at the ER membrane, thereby controlling TA protein biogenesis.

Description: This study investigates pressure effects on the magnetic properties of non-interacting single-domain (SD) magnetite. Using a high-pressure cell specially designed for a Magnetic Property Measurement System, magnetic hysteresis measurements were conducted under high pressures of up to 1 GPa on natural plagioclase crystals containing much acicular SD magnetite. Coercivity and saturation magnetization were nearly constant with pressure, while saturation remanent magnetization and coercivity of remanence decreased with pressure at moderate rates of –8 per cent GPa –1 and –18 per cent GPa –1 , respectively. These results suggest that temperature effects govern the magnetic behaviour of acicular SD magnetite grains in the middle and lower crusts.

Description: The ultrastructure of melanin granules in human hair was studied using 1,000 kV high-voltage scanning transmission electron microscopy to successfully reconstruct three-dimensional images of the whole melanin granule. It was revealed that the melanin granule was composed of a membrane-like outer structure that included many spherical vesicles, and an inner matrix containing a sheet-like structure in the elongated direction of the melanin granule and a sheet-like arrays structure in the cross direction. The outer structure of the melanin granule was maintained even after exposure to hair-bleaching agents to decompose the melanin granule, suggesting that the outer structure was a highly robust structure and composition compared with the inner matrix .

Description: Palaeointensity experiments were carried out to a sample collection from two sections of basalt lava flow sequences of Pliocene age in north central Iceland (Chron C2An) to further refine the knowledge of the behaviour of the palaeomagnetic field. Selection of samples was mainly based on their stability of remanence to thermal demagnetization as well as good reversibility in variations of magnetic susceptibility and saturation magnetization with temperature, which would indicate the presence of magnetite as a product of deuteric oxidation of titanomagnetite. Among 167 lava flows from two sections, 44 flows were selected for the Königsberger–Thellier–Thellier experiment in vacuum. In spite of careful pre-selection of samples, an Arai plot with two linear segments, or a concave-up appearance, was often encountered during the experiments. This non-ideal behaviour was probably caused by an irreversible change in the domain state of the magnetic grains of the pseudo-single-domain (PSD) range. This is assumed because an ideal linear plot was obtained in the second run of the palaeointensity experiment in which a laboratory thermoremanence acquired after the final step of the first run was used as a natural remanence. This experiment was conducted on six selected samples, and no clear difference between the magnetic grains of the experimented and pristine sister samples was found by scanning electron microscope and hysteresis measurements, that is, no occurrence of notable chemical/mineralogical alteration, suggesting that no change in the grain size distribution had occurred. Hence, the two-segment Arai plot was not caused by the reversible multidomain/PSD effect in which the curvature of the Arai plot is dependent on the grain size. Considering that the irreversible change in domain state must have affected data points at not only high temperatures but also low temperatures, f v ≥ 0.5 was adopted as one of the acceptance criteria where f v is a vectorially defined fraction of the linear segment. A measure of curvature k ' was also used to check the linearity of the selected linear segment. It was avoided, however, to reject the result out of hand by the large curvature k of the entire data points because it might still include a linear segment with a large fraction. Combining with the results of Shaw's experiments, 52 palaeointensities were obtained out of 192 specimens, or 11 flow means were obtained out of the 44 lava flows. Most of the palaeointensities were from the upper part of the lava section (Chron C2An.1n) and ranged between 30 and 66 μT. Including two results from the bottom part of the lava section, the mean virtual dipole moment for 2.5–3.5 Ma is 6.3 ± 1.4  x  10 22 Am 2 ( N  = 11), which is ~19 per cent smaller than the present-day dipole moment.

Description: In this article, we propose a smart image-analysis method suitable for extracting target features with hierarchical dimension from original data. The method was applied to three-dimensional volume data of an all-solid lithium-ion battery obtained by the automated sequential sample milling and imaging process using a focused ion beam/scanning electron microscope to investigate the spatial configuration of voids inside the battery. To automatically fully extract the shape and location of the voids, three types of filters were consecutively applied: a median blur filter to extract relatively larger voids, a morphological opening operation filter for small dot-shaped voids and a morphological closing operation filter for small voids with concave contrasts. Three data cubes separately processed by the above-mentioned filters were integrated by a union operation to the final unified volume data, which confirmed the correct extraction of the voids over the entire dimension contained in the original data.

Description: Cation mixing in positive electrode materials for rechargeable lithium ion batteries, LiNi x Mn 2– x O 4 ( x = 0, 0.2, 0.5) and Li 0.21 Ni 0.7 Mn 1.64 O 4– (denoted as x = 0.7), is analyzed by high-angular-resolution electron-channeling X-ray/electron spectroscopy (HARECXS/HARECES) techniques, using energy-dispersive X-ray spectroscopy and electron energy-loss spectroscopy. Mixing between the tetrahedral lithium sites and the octahedral transition metal sites is quantified, and the site-dependent valence states of the transition metals are examined. In the non-doped ( x = 0) sample, Mn was found to occupy only octahedral sites as either Mn 3+ or Mn 4+ . For x = 0.2–0.7, some of the nickel ions (6–13% depending on x ) occupy tetrahedral anti-sites. All the nickel ions are in the divalent state, regardless of the occupation site. For x = 0.2 and 0.7, manganese ions occupy both octahedral and tetrahedral sites; those in the octahedral sites are tetravalent, while the tetrahedral sites contain a mixture of divalent and trivalent ions. For x = 0.5, manganese occupies only the octahedral sites, with all ions determined to be in the tetravalent state (within experimental accuracy). All the samples substantially satisfied the local charge neutrality conditions. This study demonstrates the feasibility of using HARECXS/HARECES for quantitative analysis of the atomic configuration and valence states in lithium manganese oxide spinel materials.

Description: Seipin, encoded by BSCL2 gene, is a protein whose physiological functions remain unclear. Mutations of BSCL2 cause the most-severe form of congenital generalized lipodystrophy (CGL). BSCL2 mRNA is highly expressed in the brain and testis in addition to the adipose tissue in human, suggesting physiological roles of seipin in non-adipose tissues. Since we found BSCL2 mRNA expression pattern among organs in rat is similar to human while it is not highly expressed in mouse brain, we generated a Bscl2 /seipin knockout (SKO) rat using the method with ENU ( N -ethyl- N -nitrosourea) mutagenesis. SKO rats showed total lack of white adipose tissues including mechanical fat such as bone marrow and retro-orbital fats, while physiologically functional brown adipose tissue was preserved. Besides the lipodystrophic phenotypes, SKO rats showed impairment of spatial working memory with brain weight reduction and infertility with azoospermia. We confirmed reduction of brain volume and number of sperm in human patients with BSCL2 mutation. This is the first report demonstrating that seipin is necessary for normal brain development and spermatogenesis in addition to white adipose tissue development.

Description: Hibernation-specific protein (HP) is a plasma protein that regulates hibernation in chipmunks. The HP complex (HP20c) consists of three homologous proteins, HP20, HP25 and HP27, all produced by liver and belonging to the C1q family. To date, HP20c has not been identified in any mammalian species except chipmunk and ground squirrel hibernators. Here, we report a bovine HP20 gene isolated from liver tissue and aortic endothelial cells. Total homology between bovine and chipmunk variants was 63% at the amino acid level. Gene expression was highest in the liver. Western blot revealed HP20 protein in foetal, newborn, calf and adult serum, with highest concentrations in the adult. Similar proteins were detected in sera of other ruminants but not in humans, bears, mice or rats. Bovine HP20 protein was found mainly in ovaries, stomach, heart, kidneys, lungs, testes and prostate, but not in the skeletal muscle. Native HP20 was purified from bovine adult serum as a complex containing 25 and 27 kDa proteins. Mass spectrometry revealed that these proteins are orthologues of chipmunk HP25 and HP27, respectively. Interestingly, bovine HP20 was highly expressed in cattle serum after fasting. Native bovine HP20c may be a useful tool for investigating HP function.

Description: Structome analysis is a useful tool for identification of unknown microorganisms that cannot be cultured. In 2012, we discovered a unique deep-sea microorganism with a cell structure intermediate between those of prokaryotes and eukaryotes and described its features using freeze-substitution electron microscopy and structome analysis (quantitative and three-dimensional structural analysis of a whole cell at the electron microscopic level). We named it Myojin parakaryote . Here we describe, using serial ultrathin sectioning and high-voltage electron microscopy tomography of freeze-substituted specimens, the structome analysis and 3D reconstruction of another unique spiral bacteria, found in the deep sea off the coast of Japan. The bacteria, which is named as ‘Myojin spiral bacteria’ after the discovery location and their morphology, had a total length of 1.768 ± 0.478 µm and a total diameter of 0.445 ± 0.050 µm, and showed either clockwise or counter-clockwise spiral. The cells had a cell surface membrane, thick fibrous layer, ribosomes and inner fibrous structures (most likely DNA). They had no flagella. The bacteria had 322 ± 119 ribosomes per cell. This ribosome number is only 1.2% of that of Escherichia coli and 19.3% of Mycobacterium tuberculosis and may reflect a very slow growth rate of this organism in the deep sea.

Description: We have developed a quantitative particle size analytical method at the single atomic level employing electron microscopy and image processing for the investigation of supported metal catalysts. In the present study, a supported gold (Au) catalyst containing sub-nano clusters and individual atoms was globally observed by high-resolution high-angle annular dark-field scanning transmission electron microscopy (HAADF-STEM) using spherical aberration (Cs)-corrected TEM. To fully extract structural information of the Au clusters and individual atoms from the HAADF-STEM images, a morphological image-processing operation was applied. The resulting mean particle size was in good agreement with particle sizes estimated from average information provided by X-ray absorption fine structure analysis. It is demonstrated that the present HAADF-STEM image analysis gives a quantitative particle size distribution measurement of supported Au clusters and individual atoms.