Dynamics of cell wall formation in fission yeast, Schizosaccharomyces pombe

Studies on the dynamics of surface and intracellular structures during cell wall formation from the reverting protoplast of Schizosaccharomyces pombe were reviewed, and the correlation between cell wall formation and actin cytoskeleton, which is the most important conductor of the mechanism, is described in this paper. A close spatial and temporal relationship between actin cytoskeleton and cell wall formation was found by using wild type and actin point-mutant cps8 of S. pombe. Concomitant with the cell wall formation, dynamic behavior of the intracellular secretion machinery, especially the Golgi apparatus and secretory vesicles, was analyzed by three-dimensional reconstruction of 40 to 80 serial sections at five reverting stages. Total reverting protoplast volume increased by 3.8 and 4.3 times at 3 and 5 h, respectively, and the volume of the Golgi apparatus in the corresponding stages increased 2.3- and 2. 5-fold over the same periods. The number of secretory vesicles also markedly increased by 3.4 and 5.8 times over that of the corresponding reverting protoplasts. Actin point-mutant cps8 cells have abnormal structure in the cell wall and septum, and the distribution pattern of the actin cytoskeleton during the reversion process was different from wild-type protoplasts. The profiles of actin showed one or two thick cables and patches in the cytoplasm which remained throughout reversion. The development of crosslinkage of the glucan fibrils which are beta-1,3-glucan in nature on the reverting protoplast surface was defective; the glucan networks consisted of thin, rope-shaped fibrils up to 30 nm in width which formed a ribbon-shape 200 nm wide in wild-type reverting protoplasts. The intrafibrillar space is not filled with amorphous particles of alpha-galactomannan in nature. The secretion machinery was seen to have a similar profile as the wild type. The above results suggest that actin cytoskeleton may control secretion of beta-1,6-glucan and other cell wall substances such as alpha-glucan and alpha-galactomannan rather than beta-1,3-glucan. Study of the role of actin cytoskeleton in the cell wall formation is contributing to the development of antifungal agents together with basic cell biology.     

Differences in temporal frequency tuning between the two binocular mechanisms for seeing motion in depth

There are two types of binocular cues available for perception of motion in depth. One is the binocular disparity change in time and the other is the velocity difference between the left and the right retinal images (inter-ocular velocity differences). We measured the luminance contrast threshold for seeing motion in depth while isolating either of the cues at various temporal modulations of velocity in the stimulus. To isolate disparity cues, dynamic random-dot stereograms were used (the disparity condition) while binocularly uncorrelated random-dot kinematograms were used to isolate velocity cues (the velocity condition). Results showed that sensitivity peaked at a temporal frequency (approximately 1 cps) in the velocity condition while the peak in the disparity condition was at the lowest frequency (0.35 cps) or at least at a frequency lower than that in the velocity condition. This suggests that the visual system has different temporal frequency properties for the velocity and disparity cues for motion in depth.     

Histidine-Rich Glycoprotein Suppresses the S100A8/A9-Mediated Organotropic Metastasis of Melanoma Cells

The dissection of the complex multistep process of metastasis exposes vulnerabilities that could be exploited to prevent metastasis. To search for possible factors that favor metastatic outgrowth, we have been focusing on secretory S100A8/A9. A heterodimer complex of the S100A8 and S100A9 proteins, S100A8/A9 functions as a strong chemoattractant, growth factor, and immune suppressor, both promoting the cancer milieu at the cancer-onset site and cultivating remote, premetastatic cancer sites. We previously reported that melanoma cells show lung-tropic metastasis owing to the abundant expression of S100A8/A9 in the lung. In the present study, we addressed the question of why melanoma cells are not metastasized into the brain at significant levels in mice despite the marked induction of S100A8/A9 in the brain. We discovered the presence of plasma histidine-rich glycoprotein (HRG), a brain-metastasis suppression factor against S100A8/A9. Using S100A8/A9 as an affinity ligand, we searched for and purified the binding plasma proteins of S100A8/A9 and identified HRG as the major protein on mass spectrometric analysis. HRG prevents the binding of S100A8/A9 to the B16-BL6 melanoma cell surface via the formation of the S100A8/A9 complex. HRG also inhibited the S100A8/A9-induced migration and invasion of A375 melanoma cells. When we knocked down HRG in mice bearing skin melanoma, metastasis to both the brain and lungs was significantly enhanced. The clinical examination of plasma S100A8/A9 and HRG levels showed that lung cancer patients with brain metastasis had higher S100A8/A9 and lower HRG levels than nonmetastatic patients. These results suggest that the plasma protein HRG strongly protects the brain and lungs from the threat of melanoma metastasis.     

Tetraphenylporphyrin Enters the Ring: First Example of a Complex between Highly Bulky Porphyrins and a Protein**

Tetraphenylporphyrin (TPP) is a symmetrically substituted synthetic porphyrin whose properties can be readily modified, providing it with significant advantages over naturally occurring porphyrins. Herein, we report the first example of a stable complex between a native biomolecule, the haemoprotein HasA, and TPP as well as its derivatives. The X-ray crystal structures of nine different HasA-TPP complexes were solved at high resolutions. HasA capturing TPP derivatives was also demonstrated to inhibit growth of the opportunistic pathogen Pseudomonas aeruginosa. Mutant variants of HasA binding FeTPP were shown to possess a different mode of coordination, permitting the cyclopropanation of styrene.     

A possibility of the production of carbon nanotubes from heavy hydrocarbons

A possibility of the production of carbon nanotubes from heavy hydrocarbon resources derived from natural asphalt was examined. Before the use of heavy hydrocarbons, pure compound, toluene was used as the pure substrate to establish the reaction system for the production of carbon nanotubes. Carbon nanotubes were found in the carbonaceous product deposited on inner wall of a quartz tube and at the exit of the tube. The carbonaceous product was observed by scanning electron microscopy and analyzed by temperature-programmed oxidation experiments to identify the presence of carbon nanotubes. Based on the reaction system and reaction conditions with toluene, the production of nanotubes was examined by using heavy hydrocarbons such as asphaltene and maltene fractions from a natural asphalt. Under selected reaction conditions including the reaction temperature and the amount of the catalyst, carbon nanotubes with a diameter of 30–60 nm were found.     

A method for multidirectional TEM observation of a specific site at atomic resolution

A new technique has been developed for the three-dimensional structure characterisation of a specific site at atomic resolution. In this technique, a focused ion beam (FIB) system is used to extract a specimen from a desired site as well as to fabricate the electron transparent specimen. A specimen holder with a specimen stage rotation mechanism has also been developed for use with both an FIB system and a high-resolution transmission electron microscope (TEM). The specimen holder allows both the FIB milling of a specimen and its observation in TEM without remounting the specimen from the specimen holder. A specimen for the three-dimensional TEM observation is extracted using the FIB micro-sampling technique and shaped into a pillar to mount on a tip of a needle stub enabling a multidirectional observation. The technique was applied to the multidirectional observation of the crystal structure of an Si single crystal at atomic resolution. The crystal lattice fringes of the two Si(111) planes with distances of 0.31 nm as well as the lattice fringes of the Si(200) with distances of 0.19 nm were clearly observed.     

Room‐Temperature Printing of Organic Thin‐Film Transistors with π‐Junction Gold Nanoparticles

Printing semiconductor devices under ambient atmospheric conditions is a promising method for the large‐area, low‐cost fabrication of flexible electronic products. However, processes conducted at temperatures greater than 150 °C are typically used for printed electronics, which prevents the use of common flexible substrates because of the distortion caused by heat. The present report describes a method for the room‐temperature printing of electronics, which allows thin‐film electronic devices to be printed at room temperature without the application of heat. The development of π‐junction gold nanoparticles as the electrode material permits the room‐temperature deposition of a conductive metal layer. Room‐temperature patterning methods are also developed for the Au ink electrodes and an active organic semiconductor layer, which enables the fabrication of organic thin‐film transistors through room‐temperature printing. The transistor devices printed at room temperature exhibit average field‐effect mobilities of 7.9 and 2.5 cm² V^?1 s^?1 on plastic and paper substrates, respectively. These results suggest that this fabrication method is very promising as a core technology for low‐cost and high‐performance printed electronics.     

Towards development of enhanced fully-Lagrangian mesh-free computational methods for fluid-structure interaction

Simulation of incompressible fluid flow-elastic structure interactions is targeted by using fully-Lagrangian mesh-free computational methods. A projection-based fluid model(moving particle semi-implicit(MPS)) is coupled with either a Newtonian or a Hamiltonian Lagrangian structure model(MPS or HMPS) in a mathematically-physically consistent manner. The fluid model is founded on the solution of Navier-Stokes and continuity equations. The structure models are configured either in the framework of Newtonian mechanics on the basis of conservation of linear and angular momenta, or Hamiltonian mechanics on the basis of variational principle for incompressible elastodynamics. A set of enhanced schemes are incorporated for projection-based fluid model(Enhanced MPS), thus, the developed coupled solvers for fluid structure interaction(FSI) are referred to as Enhanced MPS-MPS and Enhanced MPS-HMPS. Besides, two smoothed particle hydrodynamics(SPH)-based FSI solvers, being developed by the authors, are considered and their potential applicability and comparable performance are briefly discussed in comparison with MPS-based FSI solvers. The SPH-based FSI solvers are established through coupling of projection-based incompressible SPH(ISPH) fluid model and SPH-based Newtonian/Hamiltonian structure models, leading to Enhanced ISPH-SPH and Enhanced ISPH-HSPH. A comparative study is carried out on the performances of the FSI solvers through a set of benchmark tests, including hydrostatic water column on an elastic plate,high speed impact of an elastic aluminum beam, hydroelastic slamming of a marine panel and dam break with elastic gate.     

Effects of mineral addition on the growth morphology of and arachidonic acid production by Mortierella alpina 1S-4

The culture conditions for arachidonic acid (AA) production by Mortierella alpina 1S-4 were investigated by means of a morphological study with the aims of obtaining a high AA yield and scale-up. In a 50-L jar fermentor study, a medium containing 3.1% soy flour and 1.8% glucose with 0.3% KH₂PO₄, 0.1% Na₂SO₄, 0.05% CaCl₂·2H₂O and 0.05% MgCl₂·6H₂O was found to be optimum. The AA yield reached 9.8 g/L/7 d, and the major morphology was small pellets (1–2 mm). However, in the case of the only addition of KH₂PO₄, the major morphology was filaments. The apparent viscosity increased to 2240 cp, thereby requiring a high agitation speed to maintain adequate oxygen tension, which caused mycelial damage due to shear stress and therefore a decrease in the AA yield. When a medium with Na₂SO₄, CaCl₂, and MgCl₂ was used, the major morphology was large pellets (2–3 mm), and mass transfer limitation through the pellet wall caused a decrease in the AA yield. Based on these results, a scale-up study was carried out under the optimal conditions described above. An AA yield of 10.9 g/L/8 d was obtained in a 10-kL industrial fermentor, and the major morphology was small pellets.     

Verification of Osakada’s method to calculate stress-strain curves from upset test load-stroke curves under various work hardening conditions

Osakada and his coworkers have suggested a method to calculate the true stress—true strain curves from the load—stroke curves obtained in the upset test which is carried out with sticking friction. The friction effect is corrected using the modification factors obtained by the finite element method (FEM). Since Osakada and his coworkers assumed no work hardening, this investigation was carried out to verify this method under various work hardening behaviors using the ALPID FEM code. It is observed that different work hardening behaviors do not affect the modification factors significantly, and thus, only one set of values can be used. A comparison with experimental results obtained with a minimum amount of friction reveals that the stress—strain curves calculated with the modification factors obtained in this investigation agree with experimental results better than those with the values Osakada suggested. Overall, it can be concluded that this modification technique is useful because it enables the analyses of the two aspects of cold forgeability, workability limit and strength, simultaneously in one upset test set-up.