One of the isoamylase isoforms, CMI294C, is required for semi-amylopectin synthesis in the rhodophyte Cyanidioschyzon merolae.

Most rhodophytes synthesize semi-amylopectin as a storage polysaccharide, whereas some species in the most primitive class (Cyanidiophyceae) make glycogen. To know the roles of isoamylases in semi-amylopectin synthesis, we investigated the effects of isoamylase gene (CMI294C and CMS197C)-deficiencies on semi-amylopectin molecular structure and starch granule morphology in Cyanidioschyzon merolae (Cyanidiophyceae). Semi-amylopectin content in a CMS197C-disruption mutant (ΔCMS197C) was not significantly different from that in the control strain, while that in a CMI294C-disruption mutant (ΔCMI294C) was much lower than those in the control strain, suggesting that CMI294C is essential for semi-amylopectin synthesis. Scanning electron microscopy showed that the ΔCMI294C strain contained smaller starch granules, while the ΔCMS197C strain had normal size, but donut-shaped granules, unlike those of the control strain. Although the chain length distribution of starch from the control strain displayed a semi-amylopectin pattern with a peak around degree of polymerization (DP) 11-13, differences in chain length profiles revealed that the ΔCMS197C strain has more short chains (DP of 3 and 4) than the control strain, while the ΔCMI294C strain has more long chains (DP ≥12). These findings suggest that CMI294C-type isoamylase, which can debranch a wide range of chains, probably plays an important role in semi-amylopectin synthesis unique in the Rhodophyta.

Cellulose intrafibrillar mineralization of biological silica in a rice plant.

The essence of morphological design has been a fascinating scientific problem with regard to understanding biological mineralization. Particularly shaped amorphous silicas (plant opals) play an important role in the vital activity in rice plants. Although various organic matters are associated with silica accumulation, their detailed functions in the shape-controlled mineralization process have not been sufficiently clarified. In the present study, cellulose nanofibers (CNFs) were found to be essential as a scaffold for silica accumulation in rice husks and leaf blades. Prior to silicification, CNFs ~ 10 nm wide are sparsely stacked in a space between the epidermal cell wall and the cuticle layer. Silica nanoparticles 20-50 nm in diameter are then deposited in the framework of the CNFs. The shape-controlled plant opals are formed through the intrafibrillar mineralization of silica nanoparticles on the CNF scaffold.

Humidity-Induced Switching between Two Magnetic and Structural Phases in a CoII -[WV (CN)8 ] Molecular Magnet.

The self-assembly of cobalt(II) with purine and octacyanidotungstate(V) results in the formation of the three-dimensional Co3 [W(CN)8 ]2 (purine)2 ⋅8.5H2 O (1) coordination polymer. This compound exhibits humidity-induced variation of the number of water molecules of crystallisation leading to a reversible structural phase transition and the alternation of the long-range ferromagnetic ordering temperature from TC =29 K for the pristine assembly (1) to TC =49 K for the sample stored in a low-humidity atmosphere (1-deh). This phenomenon can be attributed to a reversible change in the hydrogen-bonding network resulting in the modification of the local geometries of cobalt(II) as well as the cyanido bridges.

Superplasticity in an organic crystal.

Superplasticity, which enables processing on hard-to-work solids, has been recognized only in metallic solids. While metallic materials and plastics (polymer solids) essentially possess high plastic workability, functional crystalline solids present difficulties in molding. Organic crystals especially are fragile, in the common view, and they are far from the stage of materials development. From the viewpoint of practical application; however, organic crystals are especially attractive because they are composed of ubiquitous elements and often exhibit higher performance than metallic materials. Thus, finding superplastic deformation of organic crystals, especially in a single-crystal-to-single-crystal manner, will pave the way for their material applications. This study confirmed superplasticity in a crystal of a simple organic compound: N,N-dimethyl-4-nitroaniline. The crystal exhibits single-crystal-to-single-crystal superplastic deformation without heating. This finding of "organosuperplasticity" will contribute to the future design of functional solids that do not lose their crystalline quality in molding.

Electrically Activated Conductivity and White Light Emission of a Hydrocarbon Nanoring-Iodine Assembly.

Numerous otherwise difficult applications have been realized with materials, the chemical/physical properties of which can be controlled by external stimuli such as heat, pressure, photo-irradiation, and voltage bias. However, the complexity of design and the lack of easy-to-conduct synthetic methods make the creation of on-demand stimuli responsive materials a formidable task. Here we report an electric-stimuli-responsive multifunctional material, [10]CPP-I: crystalline assembly of a hydrocarbon nanoring ([10]cycloparaphenylene: [10]CPP) as an "electro-responsive porous host" and iodine as a "potentially functional molecule". Through applying electric stimulus, [10]CPP-I turned to exhibit two attractive properties: electronic conductivity and white light emission. We revealed that electric stimuli trigger the cascade formation of polyiodide chains inside the [10]CPP assembly through charge transfer, leading to the emergence of these properties. This "responsive porous host" approach is expected to be applicable for different stimuli, and opens the path for devising a generic strategy to the development of stimuli-responsive materials.

A highly porous rht-type acylamide-functionalized metal-organic framework exhibiting large CO2 uptake capabilities.

A new, highly porous acylamide-functionalized MOF with a (3,24)-connected rht-type network (HNUST-5) has been synthesized and structurally determined using powder X-ray diffraction. HNUST-5 exhibits a high BET surface area of 3643 m2 g-1, and a large CO2 uptake capability (38.9 mmol g-1 under 36 bar) with an excellent selectivity of CO2/CH4 (7.3) and CO2/N2 (32.5) at 273 K.

Differential interference contrast imaging using a pair of twisted nematic cells.

Nematic liquid crystal behaves like an optically uniaxial crystal whose optical axis coincides with the direction of molecular orientation. When an electric field is applied, a lateral shear of incident light is induced, depending on the angle of molecular inclination. While this may degrade the image quality for display applications, the precise electrical tunability of the lateral shear distance is desirable for differential interference contrast (DIC) imaging. In this Letter, a pair of twisted nematic (TN) cells is used for DIC imaging instead of the normal DIC prisms, and the unique optical properties of the TN cell are investigated for DIC imaging applications.

Cycloparaphenylene as a molecular porous carbon solid with uniform pores exhibiting adsorption-induced softness.

The molecular carbon nanoring, cycloparaphenylene (CPP), is fascinating as a new class of carbonaceous porous solids with the uniform structure of an all-benzene surface. We explored the feasibility of [12]CPP as a carbon-based porous material and uncovered its unique adsorption properties due to its shape and highly nonpolar surface. Unlike other porous carbon solids, [12]CPP shows stepwise adsorption behaviors sensitive to the functionalities of the guest molecules. In situ powder X-ray diffraction and infrared spectra provided insights into how [12]CPP accommodates the guest molecules with structural deformation retaining its structural periodicity during the whole adsorption process, which exemplifies that this molecular nanoring represents an unprecedented carbon-based soft porous solid.

External stimulation-controllable heat-storage ceramics.

Commonly available heat-storage materials cannot usually store the energy for a prolonged period. If a solid material could conserve the accumulated thermal energy, then its heat-storage application potential is considerably widened. Here we report a phase transition material that can conserve the latent heat energy in a wide temperature range, T<530 K and release the heat energy on the application of pressure. This material is stripe-type lambda-trititanium pentoxide, λ-Ti3O5, which exhibits a solid-solid phase transition to beta-trititanium pentoxide, ß-Ti3O5. The pressure for conversion is extremely small, only 600 bar (60 MPa) at ambient temperature, and the accumulated heat energy is surprisingly large (230 kJ L(-1)). Conversely, the pressure-produced beta-trititanium pentoxide transforms to lambda-trititanium pentoxide by heat, light or electric current. That is, the present system exhibits pressure-and-heat, pressure-and-light and pressure-and-current reversible phase transitions. The material may be useful for heat storage, as well as in sensor and switching memory device applications.

Determination of birefringence and slow axis distribution using an interferometric measurement system with liquid crystal phase shifter.

It is known that liquid crystal (LC) cells are useful as compact and easy-to-handle phase shifters that are readily coupled into the optics of standard microscope systems. Here, a uniformly aligned molecular LC phase shifter is introduced into a polarization microscope to attain a birefringence imaging system, using the phase-shift interferometric technique. Since the birefringence can be determined accurately only when the optical axis of the sample is parallel or perpendicular to the slow axis (variable axis) of the LC phase shifter, an improved data analysis method is proposed for determining the birefringence independently of the direction; a simple method of determining the slow axis distribution is also demonstrated. Measurements of the birefringence and slow axis distribution properties of a potato starch particle are demonstrated to confirm the novel determination method.

The Coprinopsis cinerea septin Cc.Cdc3 is involved in stipe cell elongation.

We have identified and characterized a Coprinopsis cinerea mutant defective in stipe elongation during fruiting body development. In the wild-type, stipe cells elongate at the maturation stage of fruiting, resulting in very slender cells. In the mutant, the stipe cells fail to elongate, but become rather globular at the maturation stage. We found that the mutant phenotype is rescued by a gene encoding a homolog of Saccharomyces cerevisiae CDC3 septin, Cc.Cdc3. The C. cinerea genome includes 6 septin genes, 5 of which, including Cc.cdc3, are highly transcribed during stipe elongation in the wild type. In the mutant, the level of Cc.cdc3 transcription in the stipe cells remains the same as that in the mycelium, and the level of Cc.cdc10 transcription is approximately 100 times lower than that in the wild-type stipe cells. No increase in transcription of Cc.cdc3 in the mutant may be due to the fact that the Cc.cdc3 gene has a 4-base pair insertion in its promoter and/or that the promoter region is methylated in the mutant. Overexpressed EGFP-Cc.Cdc3 fusion protein rescues the stipe elongation in the transformants, localizes to the cell cortex and assembles into abundant thin filaments in the elongating stipe cells. In contrast, in vegetative hyphae, EGFP-Cc.Cdc3 is localized to the hyphal tips of the apical cells of hyphae. Cellular defects in the mutant, combined with the localization of EGFP-Cc.Cdc3, suggest that septin filaments in the cell cortex provide the localized rigidity to the plasma membrane and allow cells to elongate cylindrically.

Intact structure of EGAM1 homeoproteins and basic amino acid residues in the common homeodomain of EGAM1 and EGAM1C contribute to their nuclear localization in mouse embryonic stem cells.

Recently, we identified the structurally related homeoproteins EGAM1, EGAM1N, and EGAM1C in both preimplantation mouse embryos and mouse embryonic stem (ES) cells. These EGAM1 homeoproteins act as positive or negative regulators of differentiation and cell growth in mouse ES cells, such that these proteins are considered transcriptional regulators. In this study, we investigated their nuclear localization and identified the amino acid residues crucial for the nuclear translocation of EGAM1 and EGAM1C. When expressed exogenously in pluripotent ES cells and somatic NIH3T3 cells, all EGAM1 homeoproteins localized to the nucleus. Analysis using the web-based tool PSORTII predicted a potential nuclear localization signal (NLS) motif, RKDLIRSWFITQRHR, in the homeodomain shared by EGAM1 and EGAM1C. The introduction of mutations, such as mutations from K or R, both basic amino acid residues, to A, in this potential NLS resulted in significant impairment of the nuclear localization of both EGAM1 and EGAM1C. In contrast, GFP fusion proteins of all the full-length EGAM1 homeoproteins failed to localize to the nucleus. These results, when taken together, suggest that basic amino acid residues in the common homeodomain of EGAM1 and EGAM1C and the intact structures of the EGAM1 homeoproteins contribute, at least in part, to the nuclear localization of these proteins in mouse ES cells.

catena-Poly[[[tetra-kis-(cyanido-κC)tungstate(IV)]-di-µ-cyanido-κC:N-bis-[diaqua-(2,2'-bipyridyl-κN,N')manganese(II)]-di-µ-cyanido-κN:C] hexa-hydrate].

The polymeric title compound, {[Mn(II) (2)W(IV)(CN)(8)(C(10)H(8)N(2))(2)(H(2)O)(4)]·6H(2)O}(n), has a one-dimensional cyanide-bridged Mn(II)-W(IV) bimetallic assembly. The coordination geometry of the W(IV) atom is eight-coordinate square-anti-prismatic and that of each of the Mn(II) atoms is six-coordinate distorted octa-hedral. Two pairs of CN ligands of W(CN)(8) are bridged to two Mn(II) atoms, the remaining CN ligands being terminal. Each Mn(II) atom is additionally coordinated by a bidentate 2,2'-bipyridyl ligand and two water mol-ecules. The crystal structure is stabilized by O-H⋯O and O-H⋯N hydrogen bonds.

Morphological and crystallographic transformation from immature to mature coccoliths, Pleurochrysis carterae.

Morphology and crystallographic orientations of coccoliths, Pleurochrysis carterae, at the various growth stages were investigated using electron back-scattered diffraction analyses and scanning electron microscope (SEM) stereo-photogrammetry to understand the developments of two different coccolith units, namely V and R units. SEM observation indicates that the immature coccolith units at the earliest stage were not perfectly fixed on the organic base plates and several units were often lacked. The all units showed platy morphology and often lay parallel to the organic base plate. Their crystal orientations were close to that of the mature R units. With further growth, the platy morphology changes to a trapezoid to anvil-shape for both units, resulting in the interlocking structure of VR units. Morphological analyses present that the edges of the platy crystals parallel to the organic base plate were estimated as <48 1>, and their inner/upper surfaces were estimated as {10 14}. As they interlocked further, R units inclined more outward to develop the inner tube elements with {10 1 4} and then each unit develops differently distal and proximal shield elements, which are respectively estimated as {10 14} in the distal view and {2 1 10} planes in the proximal view. Based on the above results, the formation of different coccolith units and their growth were discussed.

Microtexture of larval shell of oyster, Crassostrea nippona: a FIB-TEM study.

The initial formation and subsequent development of larval shells in marine bivalve, Crassostrea nippona were investigated using the FIB-TEM technique. Fourteen hours after fertilization (the trochophore stage), larvae form an incipient shell of 100-150nm thick with a columnar contrast. Selected-area electron diffraction analysis showed a single-crystal aragonite pattern with the c-axis perpendicular to the shell surface. Plan-view TEM analysis suggested that the shell contains high density of {110} twins, which are the origin of the columnar contrast in the cross-sectional images. 72h after fertilization (the veliger stage), the shell grows up to 1.2-1.4mum thick accompanying an additional granular layer between the preexisting layer and embryo to form a distinctive two-layer structure. The granular layer is also composed of aragonite crystals sharing their c-axes perpendicular to the shell surface, but the crystals are arranged with a flexible rotation around the c-axes and not restricted solely to the {110} twin relation. No evidence to suggest the existence of amorphous calcium carbonate (ACC) was found through the observation. The well-regulated crystallographic properties found in the present sample imply initial shell formation probably via a direct deposition of crystalline aragonite.

A novel highly acidic polysaccharide with inhibitory activity on calcification from the calcified scale "coccolith" of a coccolithophorid alga, Pleurochrysis haptonemofera.

Coccolith, a calcified scale with species-specific fine structure produced by marine unicellular coccolithophorid algae, consists of calcium carbonate (CaCO(3)) crystals and a small amount of organic matrices. A novel polysaccharide named coccolith matrix acidic polysaccharide (CMAP) was isolated from the coccolith of a coccolithophorid alga, Pleurochrysis haptonemofera. The structure of CMAP was determined by chemical analysis and NMR spectroscopy including COSY, TOCSY, HMQC, and HMBC to be a polysaccharide composed of the following unit: -->4) l-iduronic acid (alpha1-->2) meso-tartaric acid (3-->1) glyoxylic acid (1-->. It has four carboxyl groups per a disaccharide unit as observed in another polysaccharide PS-2 characterized previously in Pleurochrysis carterae. CMAP showed a strong inhibitory activity on CaCO(3) precipitation. These results suggest that CMAP serves as a regulator in the calcification of the coccolith.

A crustacean Ca2+-binding protein with a glutamate-rich sequence promotes CaCO3 crystallization.

The DD4 mRNA of the penaeid prawn Penaeus japonicus was shown previously to be expressed in the epidermis adjacent to the exoskeleton specifically during the post-moult period, when calcification of the exoskeleton took place. The encoded protein possessed a Ca2+-binding site, suggesting its involvement in the calcification of the exoskeleton. In the present study, an additional ORF (open reading frame) of 289 amino acids was identified at the 5' end of the previous ORF. The newly identified part of the encoded protein included a region of approx. 120 amino acids that was highly rich in glutamate residues, and contained one or more Ca2+-binding sites. In an immunohistochemical study, signals were detected within calcified regions in the endocuticular layer of the exoskeleton. Bacterially expressed partial segments of the protein induced CaCO3 crystallization in vitro. Finally, a reverse transcription-PCR study showed that the expression was limited to an early part of the post-moult period, preceding significant calcification of the exoskeleton. These observations argue for the possibility that the encoded protein, renamed crustocalcin (CCN), promotes formation of CaCO3 crystals in the exoskeleton by inducing nucleation.

Characterization of Prismalin-14, a novel matrix protein from the prismatic layer of the Japanese pearl oyster (Pinctada fucata).

The mollusc shell is a hard tissue consisting of calcium carbonate and organic matrices. The organic matrices are believed to play important roles in shell formation. In the present study, we extracted and purified a novel matrix protein, named Prismalin-14, from the acid-insoluble fraction of the prismatic layer of the shell of the Japanese pearl oyster (Pinctada fucata), and determined its whole amino acid sequence by a combination of amino acid sequence analysis and MS analysis of the intact protein and its enzymic digests. Prismalin-14 consisted of 105 amino acid residues, including PIYR repeats, a Gly/Tyr-rich region and N- and C-terminal Asp-rich regions. Prismalin-14 showed inhibitory activity on calcium carbonate precipitation and calcium-binding activity in vitro. The scanning electron microscopy images revealed that Prismalin-14 affected the crystallization of calcium carbonate in vitro. A cDNA encoding Prismalin-14 was cloned and its expression was analysed. The amino acid sequence deduced from the nucleotide sequence of Prismalin-14 cDNA was identical with that determined by peptide sequencing. Northern-blot analysis showed that a Prismalin-14 mRNA was expressed only at the mantle edge. In situ hybridization demonstrated that a Prismalin-14 mRNA was expressed strongly in the inner side of the outer fold of the mantle. These results suggest that Prismalin-14 is a framework protein that plays an important role in the regulation of calcification of the prismatic layer of the shell.

A novel calcium-binding peptide from the cuticle of the crayfish, Procambarus clarkii.

A novel peptide named calcification-associated peptide (CAP)-2 was isolated from the exoskeleton of the crayfish, Procambarus clarkii. CAP-2 consists of 65 amino acid residues and has a 44% sequence identity with CAP-1 characterized previously. It has a chitin-binding domain observed in many arthropod cuticle proteins. CAP-2 showed inhibitory activity on calcium carbonate precipitation and chitin-binding ability. A CAP-2 cDNA was cloned using RT-PCR and RACE and the open reading frame encoded a precursor peptide consisting of a signal peptide and CAP-2. RT-PCR revealed that CAP-2 mRNA was exclusively expressed in the epidermal tissue during the postmolt stage, the site and stage being associated with calcification. Calcium-binding assay using recombinant CAP-2 revealed that this peptide had affinity for calcium ions with a Kd value of about 1 mM. All these results suggest that CAP-2 serves as a nucleator or a regulator in the calcification of the exoskeleton.

Cloning and expression of a cDNA encoding a matrix peptide associated with calcification in the exoskeleton of the crayfish.

Calcification-associated peptide (CAP)-1 isolated from the exoskeleton of the crayfish, Procambarus clarkii, has anti-calcification activity and chitin-binding ability and is, therefore, considered to be associated with calcification. In this study, a cDNA encoding CAP-1 was cloned and characterized. An open reading frame encoded a pre-propeptide of 99 amino acid residues, which was composed of a signal peptide, a CAP-1 precursor and two-basic amino acid residues at the C-terminus. The dibasic residues were not observed in the natural CAP-1. Expression analyses using Northern blot and RT-PCR revealed that the mRNA encoding CAP-1 was strongly expressed in the epidermal tissue during the postmolt stage, where and when the calcification takes place. These results support that CAP-1 may play an important role in the calcification of the exoskeleton. Based on the nucleotide sequence of the cDNA encoding CAP-1, a recombinant CAP-1 and that carrying the basic residues at the C-terminus were expressed in Escherichia coli. Anti-calcification assay showed that these recombinant peptides were less active than natural CAP-1, indicating that the phosphate group at the 70th residue, Ser, in natural CAP-1 is important for inhibitory activity and that the paired basic residues have some contribution to the elevation of inhibitory activity.