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Glycoprotein non-metastatic melanoma protein B (GPNMB) is a transmembrane protein enriched on the cell surface of cancer cells, including melanoma, glioblastoma, and triple-negative breast cancer. There is growing evidence identifying GPNMB as a tumor-promoter; however, despite its biological and clinical significance, the molecular mechanisms engaged by GPNMB to promote tumorigenesis are not well understood. GPNMB promotes aggressive behaviors such as tumor cell proliferation, migration, and invasion. The extracellular domain of GPNMB shed from the cell surface interacts with integrins to facilitate in the recruitment of immune-suppressive and pro-angiogenic cells to the tumor microenvironment, thereby enhancing tumor migration and invasion. GPNMB also modulates receptor tyrosine kinases and integrin signaling in a cell autonomous fashion, leading to downstream kinase signaling that in turn triggers the expression and secretion of tumorigenic factors such as matrix metalloproteinases (MMPs) and cytokines. Therefore, GPNMB exerts its pro-tumorigenic role both intracellularly and in a paracrine fashion through shedding its extracellular domain. This review highlights the importance of GPNMB in cancer progression and discusses molecular mediators of GPNMB-induced tumor growth and invasion., (Copyright © 2017. Published by Elsevier Inc.)

The utility of engineering flocculation is wildly recognized in applied and environmental microbiology. We previously reported self-produced flocculation of Escherichia coli cells by overexpressing the native bcsB gene that encodes a component of the cellulose synthesis pathway. Further experiments clarified that the spontaneous E. coli flocs were proteinous, and elongation factor Ts (Tsf) was the main component. In this study, we demonstrated successful expression of a fusion protein consisting of Tsf and green fluorescence protein (GFP) on E. coli flocs. Interestingly, the percentage of Tsf-GFP in total floc protein reached approximately 15% (w/w). The proposed design of a fusion protein with Tsf enables displaying a recombinant target protein on the floc structure., (Copyright © 2017 Elsevier Inc. All rights reserved.)

Hydrogel fibers with biodegradable and biocompatible features are useful for the fabrication of filament-like tissues. We developed cell-laden hyaluronic acid (HA)-based hollow hydrogel fibers to create single and bundled filament-like tissues. The cell-laden fibers were fabricated by crosslinking phenolic-substituted hyaluronic acid (HA-Ph) in an aqueous solution containing cells through a horseradish peroxidase (HRP)-catalyzed reaction in the presence of catalase by extruding the solution in ambient flow of an aqueous solution containing H 2 O 2 . The encapsulated cells proliferated and grew within the hollow core, and the cells formed filament-like constructs in both single and bundled fibers, which were obtained by collection on a rotating cylindrical tube. Single and bundled filament-like tissues covered with an additional heterogeneous cell layer were obtained by degrading the fiber membrane using hyaluronidase after covering the fiber surface with heterogeneous cells. Cellular viability was preserved during HA-Ph hydrogel fiber fabrication and filament-like tissue formation. These results demonstrate the feasibility of HA-based hollow hydrogel fibers obtained through HRP- and catalase-mediated reactions to engineer filament-like tissues., (Copyright © 2017 Elsevier B.V. All rights reserved.)

Biological materials are the result of years of evolution and possess a number of efficient features and structures. Researchers have investigated the possibility of designing biomedical structures that take advantage of these structural features. Insect shells, such as beetle shells, are among the most promising types of biological material for biomimetic development. However, due to their intricate geometries and small sizes, it is challenging to measure the mechanical properties of these microscale structures. In this study, we developed an in-situ testing platform for site-specific experiments in a focused ion beam (FIB) system. Multi-axis nano-manipulators and a micro-force sensor were utilized in the testing platform to allow better results in the sample preparation and data acquisition. The entire test protocol, consisting of locating sample, ion beam milling and micro-mechanical bending tests, can be carried out without sample transfer or reattachment. We used our newly devised test platform to evaluate the micromechanical properties and structural features of each separated layer of the beetle horn shell. The Young's modulus of both the exocuticle and endocuticle layers was measured. We carried out a bending test to characterize the layers mechanically. The exocuticle layer bent in a brick-like manner, while the endocuticle layer exhibited a crack blunting effect., Statement of Significance: This paper proposed an in-situ manipulation/test method in focused ion beam for characterizing micromechanical properties of beetle horn shell. The challenge in precise and accurate fabrication for the samples with complex geometry was overcome by using nano-manipulators having multi-degree of freedom and a micro-gripper. With the aid of this specially designed test platform, bending tests were carried out on cantilever-shaped samples prepared by focused ion beam milling. Structural differences between exocuticle and endocuticle layers of beetle horn shell were explored and the results provided insight into the structural advantages of each biocomposite structure., (Copyright © 2017 Acta Materialia Inc. Published by Elsevier Ltd. All rights reserved.)

The hydrogels having the ability to promote migration and morphogenesis of endothelial cells (ECs) are useful for fabricating vascularized dense tissues in vitro. The present study explores the immobilization of low molecular weight hyaluronic acid (LMWHA) derivative within gelatin-based hydrogel to stimulate migration of ECs. The LMWHA derivative possessing phenolic hydroxyl moieties (LMWHA-Ph) was bound to gelatin-based derivative hydrogel through the horseradish peroxidase-catalyzed reaction. The motility of ECs was analyzed by scratch migration assay and microparticle-based cell migration assay. The incorporated LMWHA-Ph molecules within hydrogel was found to be preserved stably through covalent bonds during incubation. The free and immobilized LMWHA-Ph did not lose an inherent stimulatory effect on human umbilical vein endothelial cells (HUVECs). The immobilized LMWHA-Ph within gelatin-based hydrogel induced the high motility of HUVECs, accompanied by robust cytoskeleton extension, and cell subpopulation expressing CD44 cell receptor. In the presence of immobilized LMWHA-Ph, the migration distance and the number of existing HUVECs were demonstrated to be encouraged in dose-dependent and time-dependent manners. Based on the results obtained in this work, it was concluded that the enzymatic immobilization of LMWHA-Ph within gelatin-based hydrogel represents a promising approach to promote ECs' motility and further exploitation for vascular tissue engineering applications., (Copyright © 2017 Elsevier B.V. All rights reserved.)

Polyethylene glycol (PEG) derivatives possessing oleyl and reactive groups for conjugating functional substrates, such as proteins and quantum dots, are useful materials for cell-surface engineering and cell immobilization onto substrates. The reagent is known as a biocompatible anchor for cell membranes (BAM). Here, BAM-anchoring on cell membranes is reported to stimulate reactive oxygen species (ROS) production in those cells. Significant increases in ROS production and release to the surrounding environment were detected in mouse fibroblast cell line 10T1/2 when soaked in a solution containing BAM conjugated with 1/10mol/mol bovine serum albumin at 1.5μM-protein. ROS production stimulation was confirmed to be independent of the protein crosslinked with BAM and of cell type. Similar stimulation was detected for BAMs conjugated with ovalbumin and casein, in human hepatoma cell line HepG2, and human umbilical vein endothelial cells. Considering the effects of ROS on a variety of cellular processes, these results demonstrated the necessity for focusing attention on the effects of generated and released ROS on the behaviors of cells in the studies applying BAM to cells., (Copyright © 2016 Elsevier B.V. All rights reserved.)

The influence of contents of galactose and phenolic hydroxyl (Ph) groups incorporated into chitosan was investigated on characteristics of the chitosan derivatives and the resultant gels as well as HepG2 cell attachment and growth behaviors. Introduction of galactose groups increased the solubility of the chitosan derivatives. The gelation time decreased with increasing content of Ph groups in the chitosan derivatives. The increase of galactose groups incorporated at a fixed content of Ph groups improved mechanical properties of the resultant gels. In vitro degradation rate of the resultant gels decreased by increasing Ph groups and decreasing galactose groups incorporated into the chitosan derivatives. The HepG2 cells formed dense spheroid cell clusters when the galactose groups were absent or incorporated at high level into chitosan (13.8mol%). However, the cells exhibited spreading morphology with spheroid formation on the gels containing 1.1 and 5.2mol% galactose groups. The albumin secretion level on a cellular basis also increased considerably when the galactose groups increased to 13.8mol%. The results demonstrated the potential of the chitosan derivative hydrogels for liver tissue engineering applications., (Copyright © 2016 Elsevier B.V. All rights reserved.)

Postsurgical peritoneal adhesion is a serious surgical complication. In situ hydrogel formation on the surface of tissues, which will develop adhesions, is a recent feasible approach to prevent peritoneal adhesion. Here, we report on-tissue surface formation of a hyaluronic acid-based hydrogel by administration of a pre-hydrogel aqueous solution. The hydrogelation was initiated by contact with body fluid containing glucose on tissue surfaces. During the hydrogelation, a hyaluronic acid derivative possessing phenolic hydroxyl moieties (HA-Ph) was cross-linked by a cascade reaction of glucose oxidase (GOx) and horseradish peroxidase (HRP). About 5s of hydrogelation was accomplished using a solution containing 1.5% (w/v) HA-Ph, 5U/mL HRP, and 2.5U/mL GOx in 1mg/mL glucose that is equivalent to the normal blood glucose concentration. The hydrogel was degradable by hyaluronidase and much softer than rat peritoneal sidewalls. We confirmed the efficiency of the hydrogel to prevent post-operative peritoneal adhesions by applying the solution containing HA-Ph, GOx, and HRP to animals with bowel abrasion-abdominal sidewall defects. A significant reduction in the development of peritoneal adhesions was found compared with animals applied with phosphate-buffered saline or saline containing HA-Ph alone., Statement of Significance: Postsurgical peritoneal adhesion is a serious surgical complication. In this paper, we report a novel system for preventing it through an on-tissue surface formation of a biodegradable and biocompatible hyaluronic acid-based hydrogel by administration of a pre-hydrogel aqueous solution. The in situ hydrogelation is mediated by a cascade enzyme reaction of glucose oxidase (GOx) and horseradish peroxidase (HRP) initiated by contacting with body fluid containing glucose. The efficiency of the system was confirmed by applying the system to animals with bowel abrasion-abdominal sidewall defects., (Copyright © 2015 Acta Materialia Inc. Published by Elsevier Ltd. All rights reserved.)

Gelatin and alginate derivatives possessing phenolic hydroxyl moieties (gelatin-Ph and Alg-Ph) were dissolved in aqueous solution and conjugated via horseradish peroxidase-catalyzed crosslinking, resulting in hydrogelation. The objective of creating the hydrogels was to prepare cell sheets and spherical tissues wrapped in living cell sheaths. An increase in the gelatin-Ph content in the hydrogel improved cellular adhesion on the hydrogel surface but hindered degradability by alginate lyase. A hydrogel with the desired characteristics was obtained from a solution containing 0.5% (w/v) gelatin-Ph and 1.5% (w/v) Alg-Ph. Human aortic endothelial (HAE) cells and mouse embryo fibroblast 10T1/2 cells grew on the hydrogels and could be harvested as cell sheets by treatment with alginate lyase. 10T1/2 cells enclosed in Alg-Ph/gelatin-Ph microcapsules composed of the conjugate hydrogel elongated on the inner surface of the microcapsules and grew three times faster than those enclosed in Alg-Ph microcapsules. Alg-Ph/gelatin-Ph microcapsules not only supported growth of the enclosed cells into spherical tissues, but also provided a cell adhesive outer surface for the fabrication of an HAE cell layer. Finally, spherical tissues of 10T1/2 cells wrapped in living HAE cell sheaths were obtained by treatment with alginate lyase., (Copyright © 2013 Acta Materialia Inc. Published by Elsevier Ltd. All rights reserved.)

The phase correction function used in the three-dimensional Fourier filtering method (3D-FFM) for compensating lens aberrations was investigated to reconstruct a high-resolution exit wave of a sample. An appropriate function, which hardly suffered from imperfect illumination conditions, was determined by comparing two types of phase correction functions with numerical calculations and experiments using through-focus images of an amorphous thin film and a [110]-oriented Si single crystal taken under tilted illumination or partially coherent illumination. Theoretical calculations indicated that a function in terms of w (an axial Fourier component), available uniquely in the 3D Fourier space, compensated for the phase shift due to the spherical aberration more precisely than did a conventional function in terms of g (the two-dimensional (2D) planar Fourier components). Experimentally, exit waves reconstructed using the w-function showed sample structures at approximately 20% higher resolution than those reconstructed using the g-function. Image contrast simulations proved that the w-function had a significant advantage over the g-function: the former canceled out the effect of illumination divergence, resulting in a high-resolution exit wave. These results demonstrated that exit waves, which are uniquely realized in the 3D-FFM, should be reconstructed using the w-type phase correction function.

In order to characterize the contributions of respiratory and photosynthetic actions to energy conversions, the mixotrophic cells of Marchantia polymorpha were cultivated in the medium containing 10kg/m(3) glucose as an organic carbon source. The cultures were conducted with the supply of ordinary air (0.03% CO(2)) at constant incident light intensities of 50 and 180W/m(2). From the results of metabolic analysis, it was found that the cell yield based on ATP synthesis was estimated to be 6.3x10(-3)kg-dry cells/mol-ATP in these cultures. Under the examined conditions, energy conversion efficiency through respiration was larger than that through photosynthesis, and efficiency of overall energy conversion to ATP was maximized when the sum of energies from glucose and light captured by the cells was approximately 7.2x10(5)J/(hkg-dry cells). Taking into account the efficiency of overall energy conversion, a batch culture of M. polymorpha in a bioreactor was carried out by regulating incident light intensity ranging from 9 to 58W/m(2). In the culture with light regulation, the cell yield of 6.2x10(-9)kg-dry cells/J was achieved on the basis of energy provided to the system throughout the culture, and this value was 2.3 and 9.3 times as large as those obtained in the cultures under constant incident light intensities of 50 and 180W/m(2), respectively.

The effect of sucrose concentration in the medium on the growth and resumption ability to form lateral roots was investigated using the hairy roots of pak-bung and tobacco. It was found that the growth evaluated by root tip elongation of pak-bung and tobacco hairy roots was suppressed in the medium having an initial sucrose concentration of <2.5kg/m(3), and that the resumption abilities of both the hairy roots could be preserved when the hairy roots were kept at an initial sucrose concentration of 2.5kg/m(3) under ambient temperature conditions. The values of maintenance energy for pak-bung and tobacco hairy roots were determined to be 0.11 and 0.12 per day, respectively, from the total sugar consumption rates. Under the oligotrophic condition of the sucrose concentration of 2.5kg/m(3), the hairy roots were considered to exist as resting cells with maintenance metabolism, and the minimum demand for the energy source to ensure survival of the cells was met because the cells hardly multiplied and sugar consumption was not significant. In addition, long-term storage of pak-bung hairy roots in the liquid medium with 2.5kg/m(3) sucrose was performed at 25 degrees C. It was demonstrated that the hairy roots could maintain their resumption abilities without a serious loss of viability over 600 days and that the number of budding lateral roots per unit length of the main roots remained a value of 72 roots/m after the 600-day storage.

In the culture of red beet hairy roots in shaking flasks, a period for acclimation without lateral root generation existed at the early stage, and the root tip meristems containing growing points (GPs) were found to be damaged under an elevated shear stress condition. The loading experiments of shear stress to the hairy roots revealed that the GPs subjected to the acclimation acquired tolerance to shear stress, retaining relatively high viability of GPs up to 0.6N/m(2) of loaded shear stress. Next, the hairy roots after culture for 50h at 0.05N/m(2) of shear stress were exposed to conditions at various levels of shear stress in a single column reactor, and a relatively high growth rate was obtained in the vicinity of 1.0N/m(2) of shear stress. According to these results, two-stage cultures of hairy roots were then performed, which was comprised of a first stage for 50h at 0.05N/m(2) of shear stress for the prevention of decay of the GPs caused by hydraulic stress and a second stage for 110h at 1.0N/m(2) of shear stress for active elongation of the GPs with sufficient nutrient supply by regulation of the medium flow rate. The cell concentration ultimately reached 7.6kg dry cells/m(3), although no growth was observed in the case where the hairy roots did not undergo the first stage.

The purpose of this study was to clarify the clinical significance of rest-redistribution in myocardial scintigraphy (SPECT) in acute myocardial infarction (AMI). Thirty patients with AMI within one week after the onset of attack were studied. SPECT images were obtained 10 min and three hours after injection of 201Tl. Bull's eye images and unfolded map images were prepared. A 201Tl uptake was studied at the infarct and non-infarct sites. Exercise SPECT and radionuclide angiography (RNA) were performed in all patients one month after the onset of AMI, and the findings were compared with clinical and coronary angiographic (CAG) findings. 1. Redistribution of thallium at rest was observed at the infarct sites in nine of the 30 patients. 2. Redistribution at rest was observed at the non-infarct sites in eight patients. 3. Redistribution at rest was observed during exercise SPECT one month after the onset of AMI in patients with redistribution at rest in the acute phase. 4. In patients with redistribution at rest at the infarct site, left ventricular ejection fraction (EF) improved one month after the onset of AMI (delta EF greater than 5%), but it decreased slightly during exercise. 5. Wall motion at the infarct site was not much impaired in patients who showed redistribution at rest at the infarct site. 6. Angina pectoris and recurrence of myocardial infarction were observed more frequently in those with redistribution at rest on SPECT. 7. No characteristic findings were obtained on CAG in those with redistribution at rest.(ABSTRACT TRUNCATED AT 250 WORDS)