Dynamics and biogeochemical significance of the physical environment in Lake Biwa

We have examined two aspects of the dynamics and biogeochemical significance of the physical environment in Lake Biwa. One is the horizontal distribution of cyanobacteria as it relates to the ‘first gyre’ in the north basin of Lake Biwa. We could easily measure the first gyre using a vessel‐mounted acoustic doppler currents profiler. We were able to quantify the dynamics of the horizontal and vertical structure of currents and water temperature of this gyre. The first gyre did not remain at a fixed position; it moved north and south according to the growth of the gyre. This may play a role in the redistribution of cyanobacteria from place to place in the north basin. The second important environmental dynamic we measured was oxygen consumption rates from 1994 to 2000. We found that the minimum oxygen concentration in the hypolimnion has a clear inverse relationship with the apparent oxygen consumption rates in the 80–85 m depth layer. As a reduction in oxygen concentration in the hypolimnion can have a serious impact on benthic organisms, we concluded that the Lake Biwa environment should be monitored carefully and systematically.     

Improvement of lysozyme recovery method from the precipitate of protein-anionic surfactant complexes

A new protein separation process using a surfactant and a polar organic solvent consists of a precipitation step and a recovery step. In the precipitation step, a protein-surfactant complex is precipitated from an aqueous solution, when an ionic surfactant, sodium di(2-ethylhexyl) sulfosuccinate (AOT), is added to an aqueous solution, including protein (lysozyme). In the recovery step, the precipitate is dissolved in a polar organic solvent, such as acetone, and the protein is recovered as precipitates when a very small amount of salt solution was added to remove surfactants from the protein-surfactant complex. However, the details of the protein recovery step from precipitate have not been studied yet. In this study, the improvement of the protein recovery step was examined from the viewpoint of a recovery ratio of protein and a remaining ratio of surfactant. The optimum NaCl concentration in the feed for the protein recovery was in the range of 0.05–0.2 kmol/m³. As the NaCl concentration in the feed increased to more than 0.2 kmol/m³, the precipitation ratio decreased due to the electrostatic screening effect of NaCl. It was found that the addition of a very small amount of NaCl solution to acetone was unnecessary when NaCl was included in the feed lysozyme solution. On the other hand, as the NaCl concentration decreased to less than 0.05 kmol/m³, the precipitation ratio was decreased due to the low re-precipitation of protein by the addition of a small amount of NaCl solution in acetone. In the case of the feed containing no salt, the desired NaCl concentration added to acetone was in the range above 0.2 kmol/m³. In addition, the most suitable volume ratio of acetone to feed was found to be 0.2.     

Biosynthetic Short Neuropeptides: A Rational Theory Based on Experimental Results for the Missing Pain-Relief Opioid Endomorphin Precursor Gene

Endomorphins are neuropeptides that bind strongly to μ-opioid receptors and are considered to play important roles in pain modulation and other biological functions. Two endomorphins have been identified, to date, endomorphine-1 and -2; both are tetrapeptides and differ by only a single amino acid in the third position. Both peptides were isolated from bovine brains; however, their precursor genes have not been identified. In this study, a nucleotide sequence corresponding to the endomorphin-1 peptide in an expressed sequence tag database has been found and a preproendomorphin-like precursor peptide from human brain complementary DNA (cDNA) has been cloned. The cDNA consists of nucleotide sequences of two already annotated predicted genes, and the putative peptide differs by one amino acid from the isolated endomorphin peptides. It is proposed herein that there is the possibility of unknown short proteins or peptide precursors being missed by automated gene prediction programs based on similarities of known protein sequences. A novel concept of how to produce endomorphins from a similar peptide is described. The oxidatively modified base might provide a clue for understanding discrepancies between nucleotide sequences on the genome and those on cDNAs.     

Efficient and precise cutting of zirconia ceramics using heated cutting tool

The thermally assisted machining of yttria-stabilized tetragonal zirconia polycrystal using a cutting tool heated with induction heating was proposed. Although the conventional thermally assisted machining cannot be applied to drilling, the proposed method can be. Heat transfer from the heated cutting tool to the workpiece was simulated analytically, and the result showed that heating of the tool up to 500 °C produced an increase of 150–400 °C in the workpiece temperature. Cutting experiments demonstrated an improvement in machinability.     

Identification of Neoantigens in Cancer Cells as Targets for Immunotherapy

The clinical benefits of immune checkpoint blockage (ICB) therapy have been widely reported. In patients with cancer, researchers have demonstrated the clinical potential of antitumor cytotoxic T cells that can be reinvigorated or enhanced by ICB. Compared to self-antigens, neoantigens derived from tumor somatic mutations are believed to be ideal immune targets in tumors. Candidate tumor neoantigens can be identified through immunogenomic or immunopeptidomic approaches. Identification of neoantigens has revealed several points of the clinical relevance. For instance, tumor mutation burden (TMB) may be an indicator of immunotherapy. In various cancers, mutation rates accompanying neoantigen loads may be indicative of immunotherapy. Furthermore, mismatch repair-deficient tumors can be eradicated by T cells in ICB treatment. Hence, immunotherapies using vaccines or adoptive T-cell transfer targeting neoantigens are potential innovative strategies. However, significant efforts are required to identify the optimal epitopes. In this review, we summarize the recent progress in the identification of neoantigens and discussed preclinical and clinical studies based on neoantigens. We also discuss the issues remaining to be addressed before clinical applications of these new therapeutic strategies can be materialized.     

Model-less visual servoing using modified simplex optimization

This article describes a robot positioning task with respect to a static target by visual servoing. The vision system is uncalibrated, and the kinematic model of the robot may be totally unknown. The displacements of the robot at joint level are generated in real time in order to minimize the objective function. The objective function includes the quadratic error between the current and the desired target images. A simplex method is used to minimize the objective function, and a Newton-like method is also used near convergence. We successfully validated this method with simulations under the graphic library OpenGL. This work was presented in part at the 10th International Symposium on Artificial Life and Robotics, Oita, Japan, February 4–6, 2005     

Albumin stabilizes (–)‐epigallocatechin gallate in human serum: Binding capacity and antioxidant property

(–)‐Epigallocatechin gallate (EGCg) is the major component of green tea and is known to show strong biological activity, although it can be easily oxidized under physiological conditions. In this study, we indicate that EGCg is stable in human serum and that human serum albumin (HSA) stabilizes EGCg under aerobic condition. Although EGCg is usually decomposed within 1 h in aqueous solution at neutral pH, EGCg in serum and phosphate buffer (pH 7.4) containing HSA was stable over 1 h, even at neutral and slightly alkaline pH. Under these conditions, EGCg binds to HSA non‐covalently. The sulfhydryl group acts as an antioxidant for EGCg oxidation. Incubation of EGCg with HSA is accompanied by the oxidation of a free sulfhydryl group in HSA. These results suggest that the antioxidant property and the binding capacity of HSA contribute to the stabilization of EGCg in human serum.     

Differential Effects of Fkbp4 and Fkbp5 on Regulation of the Proopiomelanocortin Gene in Murine AtT-20 Corticotroph Cells

The hypothalamic-pituitary-adrenal axis is stimulated in response to stress. When activated, it is suppressed by the negative feedback effect of glucocorticoids. Glucocorticoids directly inhibit proopiomelanocortin (Pomc) gene expression in the pituitary. Glucocorticoid signaling is mediated via glucocorticoid receptors, 11β-hydroxysteroid dehydrogenases, and the FK506-binding immunophilins, Fkbp4 and Fkbp5. Fkbp4 and Fkbp5 differentially regulate dynein interaction and nuclear translocation of the glucocorticoid receptor, resulting in modulation of the glucocorticoid action. Here, we explored the regulation of Fkbp4 and Fkbp5 genes and their proteins with dexamethasone, a major synthetic glucocorticoid drug, in murine AtT-20 corticotroph cells. To elucidate further roles of Fkbp4 and Fkbp5, we examined their effects on Pomc mRNA levels in corticotroph cells. Dexamethasone decreased Pomc mRNA levels as well as Fkpb4 mRNA levels in mouse corticotroph cells. Dexamethasone tended to decrease Fkbp4 protein levels, while it increased Fkpb5 mRNA and its protein levels. The dexamethasone-induced decreases in Pomc mRNA levels were partially canceled by Fkbp4 knockdown. Alternatively, Pomc mRNA levels were further decreased by Fkbp5 knockdown. Thus, Fkbp4 contributes to the negative feedback of glucocorticoids, and Fkbp5 reduces the efficiency of the glucocorticoid effect on Pomc gene expression in pituitary corticotroph cells.     

Adsorption of bovine serum albumin to a polymer brush prepared by atom-transfer radical polymerization in a porous inorganic membrane

Protein adsorption was performed by a polymer brush prepared by atom-transfer radical polymerization (ATRP) to a porous inorganic membrane. The porous inorganic membrane, Shirasu Porous Glass made from silica, was modified with a halogen-containing compound to bind the active species for the polymerization. Glycidyl methacrylate was polymerized from the halogen compound by ATRP for a prescribed time, and subsequently chemically modified. The progression of the chemical modification allowed the membrane to lower the phosphate-buffer flux of the porous membrane due to the attachment of the polymer brush. Bovine serum albumin (BSA), as a model protein, was adsorbed at 12 mg per gram of the membrane in permeating BSA solution through the polymer-brush-attached porous membrane.