Further Characterization of 394-GHz Gyrotron FU CW GII with Additional PID Control System for 600-MHz DNP-SSNMR Spectroscopy

A 394-GHz gyrotron, FU CW GII, has been designed at the University of Fukui, Japan, for dynamic nuclear polarization (DNP)-enhanced solid-state nuclear magnetic resonance (SSNMR) experiments at 600-MHz ¹H resonant frequency. After installation at the Institute for Protein Research (IPR), Osaka University, Japan, a PID feedback control system was equipped to regulate the electron gun heater current for stabilization of the electron beam current, which ultimately achieved stabilization of output power when operating in continuous wave (CW) mode. During exploration to further optimize operating conditions, a continuous tuning bandwidth of approximately 1 GHz was observed by varying the operating voltage at a fixed magnetic field. In the frequency range required for positive DNP enhancement, the output power was improved by increasing the magnetic field and the operating voltage from their initial operational settings. In addition, fine tuning of output frequency by varying the cavity cooling water temperature was demonstrated. These operating conditions and ancillary enhancements are expected to contribute to further enhancement of SSNMR signal.     

An essential part for Rho-associated kinase in the transcellular invasion of tumor cells

Adhesion of tumor cells to host cell layers and subsequent transcellular migration are pivotal steps in cancer invasion and metastasis. The small GTPase Rho controls cell adhesion and motility through reorganization of the actin cytoskeleton and regulation of actomyosin contractility. Cultured rat MM1 hepatoma cells migrate through a mesothelial cell monolayer in vitro in a serum-dependent, Rho-mediated manners. Among several proteins isolated as putative target molecules of Rho, the ROCK (ROK) family of Rho-associated serine-threonine protein kinases are thought to participate in the induction of focal adhesions and stress fibers in cultured cells, and to mediate calcium sensitization of smooth muscle contraction by enhancing phosphorylation of the regulatory light chain of myosin. Transfection of MM1 cells with cDNA encoding a dominant active mutant of ROCK conferred invasive activity independently of serum and Rho. In contrast, expression of a dominant negative, kinase-defective ROCK mutant substantially attenuated the invasive phenotype. A specific ROCK inhibitor (Y-27632) blocked both Rho-mediated activation of actomyosin and invasive activity of these cells. Furthermore, continuous delivery of this inhibitor using osmotic pumps considerably reduced the dissemination of MM1 cells implanted into the peritoneal cavity of syngeneic rats. These results indicate that ROCK plays an essential part in tumor cell invasion, and demonstrate its potential as a therapeutic target for the prevention of cancer invasion and metastasis.     

Class IIa bacteriocins from lactic acid bacteria: antibacterial activity and food preservation

In the last decade, a variety of ribosomally synthesized antimicrobial peptides, or bacteriocins, produced by lactic acid bacteria have been identified and characterized. As a result of these studies, insight has been gained into various fundamental aspects of biology and biochemistry such as bacteriocin processing and secretion, mechanisms of cell immunity, and structure-function relationships. In parallel, there has been a growing awareness that bacteriocins may be developed into useful antimicrobial food additives. Class IIa bacteriocins can be considered as the major subgroup of bacteriocins from lactic acid bacteria, not only because of their large number, but also because of their significant biological activities and potential applications. The present review provides an overview of the knowledge available for class IIa bacteriocins and discusses common features and recent findings concerning these substances. The activity and potential food applications of class IIa bacteriocins are a major focus of this review.     

Suppression of Bone Necrosis around Tooth Extraction Socket in a MRONJ-like Mouse Model by E-rhBMP-2 Containing Artificial Bone Graft Administration

Medication-related osteonecrosis of the jaw (MRONJ) is related to impaired bone healing conditions in the maxillomandibular bone region as a complication of bisphosphonate intake. Although there are several hypotheses for the onset of MRONJ symptoms, one of the possible causes is the inhibition of bone turnover and blood supply leading to bone necrosis. The optimal treatment strategy for MRONJ has not been established either. BMP-2, a member of the TGF-β superfamily, is well known for regulating bone remodeling and homeostasis prenatally and postnatally. Therefore, the objectives of this study were to evaluate whether cyclophosphamide/zoledronate (CY/ZA) induces necrosis of the bone surrounding the tooth extraction socket, and to examine the therapeutic potential of BMP-2 in combination with the hard osteoinductive biomaterial, β-tricalcium phosphate (β-TCP), in the prevention and treatment of alveolar bone loss around the tooth extraction socket in MRONJ-like mice models. First, CY/ZA was intraperitoneally administered for three weeks, and alveolar bone necrosis was evaluated before and after tooth extraction. Next, the effect of BMP-2/β-TCP was investigated in both MRONJ-like prevention and treatment models. In the prevention model, CY/ZA was continuously administered for four weeks after BMP-2/β-TCP transplantation. In the treatment model, CY/ZA administration was suspended after transplantation of BMP-2/β-TCP. The results showed that CY/ZA induced a significant decrease in the number of empty lacunae, a sign of bone necrosis, in the alveolar bone around the tooth extraction socket after tooth extraction. Histological analysis showed a significant decrease in the necrotic alveolar bone around tooth extraction sockets in the BMP-2/β-TCP transplantation group compared to the non-transplanted control group in both MRONJ-like prevention and treatment models. However, bone mineral density, determined by micro-CT analysis, was significantly higher in the BMP-2/β-TCP transplanted group than in the control group in the prevention model only. These results clarified that alveolar bone necrosis around tooth extraction sockets can be induced after surgical intervention under CY/ZA administration. In addition, transplantation of BMP-2/β-TCP reduced the necrotic alveolar bone around the tooth extraction socket. Therefore, a combination of BMP-2/β-TCP could be an alternative approach for both prevention and treatment of MRONJ-like symptoms.     

Internal friction of capsule-free HIPed porous alumina

Porous alumina were sintered by conventional sintering and capsule-free Hot Isostatic Pressing (HIPing), at temperatures between 800 and 1500 °C under pressures 0.1 MPa or 200 MPa for 1 h or 50 h. Young’s modulus and internal friction of samples were measured by resonance method. The results show that Young’s modulus is mainly dominated by porosity of material. Capsule-free HIPed porous materials have slightly higher Young’s modulus than conventionally sintered ones at the same porosity. Internal friction is governed by both porosity and specific surface area. Capsule-free HIPed porous alumina has lower internal friction coefficient than conventionally sintered ones at the similar porosity or at the similar specific surface area. Enhanced surface-self diffusion under high gas pressure reduces internal friction coefficient, and affects internal friction more than Young’s modulus.     

Fabrication of highly crystalline NbOx nanotube/cup-stacked CNT nanocomposites

Carbon nanotubes (CNTs) are promising catalyst supports for fuel cell applications. Metal oxide/CNT nanocomposites are also being studied for dye-sensitized solar-cell, photocatalyst, and sensor applications. The fabrication of nanocomposites consisting of highly crystalline NbOx nanotubes and cup-stacked carbon nanotubes (CSCNTs) is reported herein. The CSCNTs were selected for the carbon materials because of their distinctive structure. The CSCNTs were photochemically treated with vacuum ultraviolet light, which increased the amount of oxygen-containing functional groups therein. NbOx nanotubes with no defects were successfully prepared with the chemical treatment of highly crystalline, layered, flux-grown K4Nb6O17 crystals. First, K4Nb6O17 crystals were grown from a KCl flux at a holding temperature of 800 degrees C. Next, NbOx nanosheets were prepared from the layered K4Nb6O17 crystals via a two-step exfoliation process, which consists of proton exchange in an acid solution and intercalation of the tetrabutylammonium ions. The NbOx nanosheets were rolled up into nanotubes with diameters of about 20 nm and lengths of 100-500 nm on the surfaces of the CSCNTs; thus, unique and complex NbOx/CSCNT nanocomposites were successfully fabricated.     

Development of an active targeting liposome encapsulated with high-density colloidal gold for transmission electron microscopy

Active targeting of the liposome is an attractive strategy for drug delivery and in vivo bio-imaging. We previously reported the specific accumulation of Sialyl Lewis X (SLX) liposome to inflamed tissue in arthritic model mice or tumor-bearing mice. SLX-liposome encapsulation with fluorescent substances allows for the visualization of these liposomes by the time-dependent transvascular accumulation of fluorescent signals in the histological sections. In the present study, we developed a new SLX-liposome encapsulated with colloidal gold for transmission electron microscopic observation. We herein describe the characterization of the colloidal gold-loaded SLX-liposomes and demonstrate its specific targeting to the endothelial cells of tumor blood vessels in tumor-bearing mice.     

Optimization of L-lactic acid feeding for the production of poly-D-3-hydroxybutyric acid by Alcaligenes eutrophus in fed-batch culture

We investigated optimization of the feeding of L-lactic acid for the production of poly-D-3-hydroxybutyric acid [P(3HB)] by Alcaligenes eutrophus in a fed-batch culture system. An acidic substrate solution was fed automatically so as to maintain the pH of the culture liquid at 7.0. Feeding of a substrate solution containing 45% (w/v) L-lactic acid, 6.2% (w/v) sodium L-lactate, 5.8% (w/v) ammonia water and 1.8% (w/v) potassium phosphate [at a molar ratio of carbon to nitrogen (C/N molar ratio) of 10], allowed the L-lactate concentration in the culture liquid to be maintained at approximately 2 g/l and the cell concentration reached 27.4 g/l after 15 h of cultivation. To promote P(3HB) production, a two-stage fed-batch culture consisting of a culture for cell growth and one for P(3HB) accumulation was carried out. When the substrate solution, whose C N molar ratio was 23, was fed during the P(3HB) accumulation phase, the cell concentration and the P(3HB) content in the cells reached 103 g/l and 57.6% (w/w), respectively, in 51.5 h.