Transient cell proliferation with polyethylenimine-cationized N-terminal domain of simian virus 40 large T-antigen

Polyethylenimine (PEI) cationization is a powerful strategy for protein transduction into cells. In this study, we attempted the artificial regulation of cell proliferation by protein transduction of the N-terminal domain (1-132 amino acids) of the simian virus 40 large T-antigen (SVLT-N), which inactivates retinoblastoma family proteins but not p53. To deliver SVLT-N into cells, we employed an indirect cationization method by forming a complex of biotynylated SVLT-N through disulfide bonds (biotin-SS-SVLT-N) and PEI-cationized avidin (PEI600-avidin). Using this complex, SVLT-N was transduced into the nucleus of confluent and quiescent Balb/c 3T3 cells and was found to be complexed with a cellular target protein, pRb. Furthermore, SVLT-N transduction induced cell proliferation in spite of confluent conditions. Because SVLT-N thus transduced into cells gradually degraded and was not detectable after a 4-d incubation, transiently transformed cells were obtained by this method. These results suggest that oncogene protein transduction technology has great potential for in vitro regulation of cell proliferation.     

Shear behaviour of reinforced concrete structural walls with eccentric openings under cyclic loading: experimental study

Reinforced concrete (RC) structural walls usually have some eccentric large openings according to the intention of the architectural design. However, the behaviour of such structures that are shear critical under cyclic loading has not been well studied. In this study, three single‐span, three‐storied, 40% scale specimens of RC structural walls with various eccentric opening ratios were constructed and tested in a lateral reverse cyclic manner until severe shear damage occurred in the walls. The main purposes of these tests were to evaluate the shear behaviour and to recognize the influence of various opening ratios on cracking and shear strength of structural walls under cyclic loading. The shear strength of specimen was calculated combining the shear strength of a structural wall without openings and the reduction factor due to openings. The comparison between the experimental results and the analytical results shows that the shear strength is different depending on the loading direction due to the eccentric opening location, the span length of short span beams may affect the cracking process and the failure mode, the approximate calculation method using Ono's reduction factor could be well applied to RC structural walls with various opening ratios less than 0.46. Copyright © 2010 John Wiley & Sons, Ltd.     

The preparation,swelling characteristics,and albumin adsorption and release behaviors of a novel chitosan-based polyampholyte hydrogel

A novel chitosan (Ch)-based polyampholyte hydrogel was prepared from Ch dissolved in a 1:1 (v/v) mixture of 10% aqueous acetic acid and N-methyl-2-pyrrolidinone (NMP) by simple crosslinking using 1,2,3,4-butanetetracarboxylic dianhydride (BTCA). The detailed structure of the hydrogel was determined via FTIR and solid-state ¹³C NMR spectroscopic analyses. The swelling behavior of the hydrogels was strongly dependent on the BTCA feed ratio, and the hydrogels exhibited a pH-responsive swelling ratio that was influenced by the presence of both cationic NH₃⁺ and anionic COO^? groups within their molecular structures. The Ch hydrogels also exhibited bovine serum albumin (BSA) adsorption capacity, which was maximal at pH 4.5, consistent with the isoelectric point of BSA (4.7). In addition, the BSA adsorption capacity of the hydrogel decreased with the increasing ionic strength of the adsorption medium, indicating that the capacity of the hydrogel to adsorb BSA is facilitated by hydrophobic as well as electrostatic interactions between the hydrogels and the BSA molecules. In addition, a high desorption ratio (89%) of BSA was achieved in aqueous solutions at pH 2.0.     

Estimation and characterisation of major royal jelly proteins obtained from the honeybee Apis merifera

Royal jelly (RJ) contains many components, including proteins. We focused on major royal jelly proteins (MRJPs) under natural conditions, and attempted to determine the content ratios and molecular forms of MRJPs by size-exclusion HPLC, SDS–PAGE, 2-DE and MALDI TOF/TOF MS. Soluble RJ proteins were extracted by dialysis followed by several centrifugation techniques. Soluble RJ proteins were universally separated into five peaks (640 kDa, 280 kDa, 100 kDa, 72 kDa and 4.5 kDa) by size-exclusion HPLC on a Superose 12 column. Among these peaks, both the 280 kDa and 72 kDa peaks were major, but the intensity of the 280 kDa peak differed markedly among original RJ samples (n = 70). The main 280 kDa protein was separated into a 55 kDa band by reducing and non-reducing SDS–PAGE. This protein was also separated into multiple spots ranging from pH 4.2 to 6.5 by 2-DE. These spots were identified as MRJP 1 by MALDI TOF/TOF MS. From these results, MRJP 1 was thought to comprise an oligomer complex linked by non-covalent bonds under natural conditions. Another major protein, the 72 kDa peak on Superose 12 HPLC, was identified as MRJP 2.     

Dynamic characteristics and dual control of a ball screw drive with integrated piezoelectric actuator

This paper describes the precision continuous path tracking control by using a dual-actuated single stage. First, fine-drive mechanism and the dynamic model of the entire drive system are described. In the simulation model, the dynamic characteristic of the dual-actuated stage is investigated to see whether it can provide precise motion by using dual control. Second, the fine motion controller is designed. Adjusting the control parameters, a positioning resolution of 20 nm and a bandwidth of 260 Hz were obtained. Third, the frequency responses of coarse and fine drives are experimentally investigated. After that, the dual controller is designed based on the investigated dynamics. Finally, whether coarse motion and fine motion could work complimentarily by the dual servo is examined in the experiments. By using the simultaneous dual controller, tracking errors were reduced sufficiently compared to the single coarse control.     

Trajectory tuning for the tracking problem of the underactuated mechanical systems: application to the PVTOL aircraft

The Planer vertical take-off and landing (PVTOL) aircraft is a typical example of an underactuated mechanical system and has a nonminimum-phase nature. When considering output tracking control, the Input/Output (I/O) linearization method is not appropriate since the stability of the internal dynamics is not guaranteed. Hauser et al. regarded this system as a slightly nonminimum-phase system which approximates to a minimum-phase one. Their control scheme yielded good results when the coupling factor was small, but the results were not acceptable when the coupling factor increased. In this article, we propose two approaches to improve the control performance. First, we consider the approximation error of Hauser's scheme as uncertainty, and apply the Linear Quadratic Regulator (LQR) method, which possesses robustness against uncertainty, to determine the stabilizing feedback coefficients. Second, from the fact that the tracking error is unavoidable, we use the “virtual reference trajectory” to design the tracking control law, and optimize this trajectory to reduce the tracking error between the “actual reference trajectory” and the “resulting trajectory”. This optimization also improves the control performance by choosing a suitable performance index. By using our approach, we achieve better performance even if the coupling factor is increased. We show these results by numerical simulation. This work was presented, in part, at the Seventh International Symposium on Artificial Life and Robotics, Oita, Japan, January 16–18, 2002     

Encrypted Thermal Printing with Regionalization Transformation

Artificially structured thermal metamaterials provide an unprecedented possibility of molding heat flow that is drastically distinct from the conventional heat diffusion in naturally conductive materials. The Laplacian nature of heat conduction makes the transformation thermotics, as a design principle for thermal metadevices, compatible with transformation optics. Various functional thermal devices, such as thermal cloaks, concentrators, and rotators, have been successfully demonstrated. How far can it possible go beyond just realizing a heat‐distribution function in a thermal metadevice? Herein, the concept of encrypted thermal printing is proposed and experimentally validated, which could conceal encrypted information under natural light and present static or dynamic messages in an infrared image. Regionalization transformation is developed for structuring thermal metamaterial‐strokes as infrared signatures, enabling letters of the alphabet to be written, paintings to be drawn, movies to be made, and information to be displayed. This strategy successfully demonstrates an extreme level of manipulation of heat flow for encryption, illusions, and messaging.     

Stabilization of linear time-invariant systems with periodic communication constraints by output sample hold control

This article considers feedback control systems wherein the control loops are closed through a real-time network, and expresses the linear time-invariant system with the constraint in an input or output as a periodic discrete time system. It is shown that this system is stabilized by using output sample hold contol. This method has the merit that the capacity of a sensor-controller communication bus is small. This work was presented, in part, at the 8th International Symposium on Artificial Life and Robotics, Oita, Japan, January 24–26, 2003     

Surface-state electrons on a hydrogen film. 1. Annealing of the film

We have investigated the surface of thin films (thickness 2 µm) of solid H₂ between 1.5 and 4.2 K by measuring the ac conductivity of surface-state electrons (SSE). The films were prepared on a glass substrate by quench condensation at 1.5 K and were therefore initially strongly disordered. In fact the surface of the virgin films before any heat treatment was so rough that no current due to SSE could be observed. Annealing the films decreased the surface roughness and gave rise to a thermal-activation-type temperature dependence of the SSE conductivity. By proper heat treatment up to 8 K the activation energy could be reduced to 10k _B.On leave from the Hyogo University of Teacher Education, Shimokume 942-1, Yashiro, 673-14, Japan.     

Phenotypic abnormalities observed in aged cloned mice from embryonic stem cells after long-term maintenance

Somatic/embryonic stem cell cloning has made it possible to produce an individual genomically identical to another individual. However, the cloned animals have a variety of abnormalities caused by the aberrant gene modification, with insufficient reprogramming in cloning. We previously reported abnormalities in cloned mice at birth. In this study, we examined what abnormalities could be seen in cloned mice after long-term maintenance. The aged cloned mice showed multiple abnormalities: increase of body weight, some phenotypic abnormalities in the kidneys, testes and thymus, and lower urea nitrogen in their serum biochemical values. The kidneys of all cloned mice were hypertrophied, with a metamorphic or whitish appearance. The multiple lesions, including the enlarged renal pelvis and distension of the renal veins in histology, might be the result of urine accumulation by urinary tract obstruction. The testes of the cloned mice were atrophied, and showed no sperm formation in histology. In contrast, the thymus was rather hypertrophied, and a comparably increased number of lymphocytes were observed in the medulla, consisting mainly of T cells. By conducting a progeny test between the cloned mice, it was confirmed that these abnormalities in the aged cloned mice were not transmitted to their offspring, indicating that the incomplete reprogramming in clones might be in part responsible for the abnormalities detected in aged clones. These results indicate that the postnatal abnormalities observed in aged cloned mice are varied and can be restored through the germ line.