Morphology of spheroidal hepatocytes within injectable, biodegradable, and thermosensitive poly(organophosphazene) hydrogel as cell delivery vehicle

An aggregate of specific cells is often regarded as a better form of single-cell in artificial organs and mammalian cell bioreactors for improved cell-specific functions. In this study, freshly harvested primary rat hepatocytes, cultivated as spheroids and entrapped in a synthetic thermoreversible extracellular matrix, were examined for differentiation morphology and enhanced liver-specific functions and compared with a control set (single-cell hepatocytes). A copolymer of poly(organophsophazene) hydrogel, an injectable, biodegradable, and thermosensitive matrix, was used to entrap hepatocytes as spheroids or single cells. In a 28-d culture period, the spheroidal hepatocytes maintained a higher viability and produced albumin and urea at constant rates, whereas the single hepatocytes showed a reduction in the level of albumin secretion in the hydrogel. The culturing of spheroidal hepatocytes is a potentially useful three-dimensional cell system for application in bioartificial liver devices and bioreactors.     

Telomerase activity and telomere lengths in various cell lines: changes of telomerase activity can be another method for chemosensitivity evaluation

For the cancer cells which have overcome the second mitotic clock (M2), activated telomerase is essential and used as another marker of immortality. Many trials had been initiated to target telomerase, which is known to be specific to tumors. To determine the best in vitro cell system for testing the efficacy of telomerase inhibitors, we evaluated the telomerase activity of various cancer cell lines and measured their telomere lengths. We also treated some cancer cell lines with adriamycin and measured the changes of telomerase activity. Telomerase activity was evaluated in various cell lines with the TRAP (telomeric repeat amplification protocol) assay. Telomerase activity was calculated and translated into arbitrary units by computer-assisted densitometry with the control of telomerase activity in the 293 control cell line. Also, terminal restriction fragment lengths were measured using Southern blotting. We also measured telomerase activity and telomere lengths in 11 benign breast tumor tissues and 19 paired stomach cancer and normal tissues. Cancer cell lines treated with adriamycin we evaluated for changes of telomerase activity and the cell proliferation by MTT assay and dye exclusion test. Telomerase activity of cell lines was 95.3 24.1 unit with a range of 27.6-129.6 unit, while the telomere lengths of those cell lines were variable from 5.0 to 10.4 kbp with a median of 6 kbp. In 11 cancer cell lines which were not yet firmly established, we could not detect any telomerase activity. Low telomerase activity was detected in only 2 benign tumor tissues of breast with a median telomere length of 8.8 (7-10.5) kbp. Among paired 19 gastric cancer and normal tissues, only 7 cancer tissues showed weak telomerase activity. After adriamycin treatment, telomerase activity in YCC-S-1, YCC-S-3, MCF-7 and MCF-7/ADR was decreased in accordance with the changes of the cell numbers. Telomerase is specific to cancer tissues and is expressed differently from organ to organ. Telomerase activity by TRAP assay could be used as a chemosensitivity assay.     

Accelerated study on lysozyme deposition on poly(HEMA) contact lenses

A technique was developed to accelerate lysozyme deposition on poly(HEMA) contact lenses and measure the amounts of the deposited lysozyme. This technique was for evaluation of bendazac lysine solution, a contact lens cleaning and wetting solution. Effect of temperature on lysozyme deposition on poly(HEMA) contact lenses was examined. Five temperatures ranging from 25 degrees C to 90 degrees C were chosen to examine the temperature effect. The amounts of lysozyme deposited on poly(HEMA) contact lenses at 25 C and 60 C were 0.27 microg/lens and 0.61 microg/lens, respectively. The amount increased sharply to 23 microg/lens at 70 degrees C with the maximum of 31 microg/lens at 90 degrees C. Kinetics of lysozyme deposition on poly(HEMA) contact lenses was examined at 80 degrees C. Lysozyme deposition increased sharply during the first 2 h and reached a plateau after 2 h. Effectiveness of various cleaning procedures was examined using bendazac lysine solution. When the contact lenses were washed without rubbing with fingers, the bendazac lysine reduced the amount of deposited lysozyme by more than 40% from 18.3 microg/lens to 10.6 microg/lens. The effect of bendazac lysine was most prominent when the contact lenses were shaken during storage in the presence of lysozyme in solution. If the contact lenses were cleaned by rubbing with fingers, the effect of bendazac lysine solution on the prevention of lysozyme deposition was negligible.     

Robust regenerative chatter stability in machine tools

The expeditious nature of manufacturing markets inspires advancements in the effectiveness, efficiency and precision of machining processes. Often, an unstable machining phenomenon, called regenerative chatter, limits the productivity and accuracies in machining operations. Since the 1950s, a substantial amount of research has been conducted on the prevention of chatter vibration in machining operations. In order to prevent regenerative chatter vibrations, the dynamics between the machine tool and workpiece are critical. Conventional regenerative chatter theories have been established based on the assumption that the system parameters in machining are constant. However, the dynamics and system parameters change due to high spindle speeds, tool geometries, orientation of the tool with respect to the rest of the machine, tool wear and non-uniform workpiece material properties. This paper provides a novel method, based on the robust stability theorem, to predict chatter-free regions for machining processes, by taking in account the unknown uncertainties and changing dynamics for machining. The effects of time-variant parameters on the stability are analyzed using the robust stability theorem. The experimental tests are performed to verify the stability of SDOF and MDOF milling systems. The uncertainties and changing dynamics are taken into account in order to accommodate the optimal selection of machining parameters, and the stability region is determined to achieve high productivity and accuracy through applications of the robust stability theorem.     

A surface wear model for hypoid gear pairs

In this paper, a methodology was proposed for simulation of surface wear of face-milled or face-hobbed hypoid gear pairs. The methodology combines Archard's wear model with a finite-element based hypoid gear contact model. The wear model requires the sliding distances and contact pressures to be computed along the contact zones at each rotational gear position. Formulations were proposed for computation of sliding distance along the tooth contact zones based on hypoid gear kinematics and geometry of the tooth surfaces, and the contact model was used to predict the normal contact pressure distributions. An example hypoid gear pair was analyzed for its wear behavior. Influences of gear position errors on wear patterns were demonstrated. An approximate method that is computationally more efficient was also proposed at the end.     

A study on reliability centered maintenance planning of a standard electric motor unit subsystem using computational techniques

The design and manufacture of urban transportation applications has been necessarily complicated in order to improve its safety. Urban transportation systems have complex structures that consist of various electric, electronic, and mechanical components, and the maintenance costs generally take up approximately 60% of the total operational costs. Therefore, it is essential to establish a maintenance plan that takes into account both safety and cost. In considering safety and cost limitations, this research introduces an advanced reliability centered maintenance (RCM) planning method using computational techniques, and applies the method to a standard electric motor unit (EMU) subsystem. First, this research devises a maintenance cost function that can reflect the current operating conditions, and maintenance characteristics, of components by generating essential cost factors. Second, a reliability growth analysis (RGA) is performed, using the Army Material Systems Analysis Activity (AMSAA) model, to estimate reliability indexes such as failure rate, and mean time between failures (MTBF), of a standard EMU subsystem, and each individual component Third, two optimization processes are performed to ascertain the optimal maintenance reliability of each component in the standard EMU subsystem. Finally, this research presents the maintenance time of each component based on the optimal maintenance reliability provided by optimization processesand reliability indexes provided by the RGA method. This paper was recommended for publication in revised form by Associate Editor Dae-Eun Kim Myung-Won Suh is a professor of Mechanical Engineering. During 1986–1988, he worked for Ford motor company as researcher. During 1989–1995, he worked in the technical center of KIA motors. He earned a BS degree in Mechanical Engineering from Seoul National University and an MS degree in Mechanical Engineering from KAIST, South Korea. He obtained his Doctorate at the University of Michigan, USA, in 1989. His research areas include structure and system optimization, advanced safety vehicle and reliability analysis & optimization. Chul-Ho Bae is a PhD candidate at Sungkyunkwan University in Suwon, South Korea. He ac-complished fellowship work as researcher at Mississippi State University, USA, in 2003 and 2005. He worked in Institute of Advanced Machinery and Technology (IMAT) as a Research Assistant in 2004. He was a part time Lecturer in computer aided Mechanical Engineering of Ansan College of Technology, Suwon Science College, and Osan College during 2004–2005. He took a BS Degree in Mechanical Design and an MS Degree in Mechanical Engineering from the Sungkyunkwan University. His research interests include computer aided engineering, reliability engineering, and optimization.     

Study on side collision reconstruction using database based on deformed shape information

A side collision reconstruction algorithm using a database based on the deformed shape information from experiments is suggested. A deformation index related to the deformed shape is developed to set the database for the side collision reconstruction algorithm. Two small-sized model cars are developed to carry out the side collision test. Several side collision tests according to velocities and collision angles are performed for establishing side collision database. A high speed camera with 1000fps is used to capture the motion of the car. Side collision reconstruction algorithm is developed and applied to find the collision conditions before the accident occurred. Several collision cases are tested to validate the database and the algorithm. A database from computer simulation is verified with experiments. According to comparing errors between simulation and experiment, it is satisfied within 6.6%. This paper was presented at the 4th Asian Conference on Multibody Dynamics(ACMD2008), Jeju, Korea, August 20–23, 2008. Dr. Jeong-Hyun Sohn received his B.S. degree from Pusan National University (1995), M.S. from PNU (1999), and Ph.D. from PNU (2002). He is currently an associate professor in the department of Mechanical Engineering at Pukyong National University in Busan, Korea. He is currently serving as a board member of dynamics and control division, KSME. His major area is vehicle dynamics and flexible multibody dynamics. Dr. Wan-Suk Yoo received his B.S. degree from Seoul National University (1976), M.S. from KAIST (1978) and Ph.D. from University of Iowa(1985). He is professor at Pusan National University, and currently serving as a vice president in KSME. His major area is vehicle dynamics and flexible multibody dynamics.     

Establishment of primary cell culture from stria vascularis explants. Morphological and functional characterization

To provide the prerequisite for long-term study of the inner ear related to structural and functional integrity, tissue of stria vascularis with spiral ligament was isolated from Wistar rat cochleas and cultured using the explant-culture technique. The following culture media were used: EMEM with Hepes buffer, hydrocortisone (400 ng/ml), transferrin (5 micrograms/ml). triiodothyronine (10(-9) M), cholera toxin (10(-10) M), insulin (5 micrograms/ml), and epidermal growth factor (10 ng/ml). To characterize the cells growing out from the explant, immunofluorescence with cytokeratin (cytokeratin 18) and ultrastructural examination with SEM and TEM were performed. The marginal cell function was investigated by expression of Na+, K(+)-ATPase antisera against beta 2 subunit of rat Na+, K(+)-ATPase and P-NPPase. We were able to maintain the cultured cells for 3 weeks or more. Monolayered marginal cells were observed beyond 14 days in vitro and the expression of cytokeratin 18 was especially enhanced. The cultured marginal cells were almost identical to in vivo cells both as regards ultrastructural features and Na+, K(+)-ATPase activity. The present results suggest that the primary explant culture technique is a reliable in vitro model of strial marginal cells. However, establishment of the cell line is needed for long-term study.     

Anatomical basis of a congenital hearing impairment: basilar papilla dysplasia in the Belgian Waterslager canary

We studied the role of proteases in apoptosis using a cell-free system prepared from a human leukemia cell line. HL60 cells are p53 null and extremely sensitive to a variety of apoptotic stimuli including DNA damage induced by the topoisomerase I inhibitor, camptothecin. We measured DNA fragmentation induced in isolated nuclei by cytosolic extracts using a filter elution assay. Cytosol from camptothecin-treated HL60 cells induced internucleosomal DNA fragmentation in nuclei from untreated cells. This fragmentation was suppressed by serine protease inhibitors. Serine proteases (trypsin, endoproteinase Glu-C, chymotrypsin A, and proteinase K) and papain by themselves induced DNA fragmentation in naive nuclei. This effect was enhanced in the presence of cytosol from untreated cells. Cysteine protease inhibitors (E-64, leupeptin, Ac-YVAD-CHO [ICE inhibitor]) did not affect camptothecin-induced DNA fragmentation. The apopain/Yama inhibitor, Ac-DEVD-CHO, and the proteasome inhibitor, MG-132, were also inactive both in the cell-free system and in whole cells. Interleukin-1 beta converting enzyme (ICE) or human immunodeficiency virus protease failed to induce DNA fragmentation in naive nuclei. Together, these results suggest that DNA damage activates serine protease(s) which in turn activate(s) nuclear endonuclease(s) during apoptosis in HL60 cells.     

A linear air bearing stage with active magnetic preloads for ultraprecise straight motion

In this paper, we propose a linear motion stage designed with magnetically preloaded air bearings. The magnetic actuators for preloading air bearings were combined with permanent magnets and coils to adjust the air bearing clearance by actively controlling the magnetic force. The system was designed to achieve a simplified configuration of air bearing stage while providing ultraprecise straight motion by actively compensating for the motion errors. The porous aerostatic bearings and magnetic preload actuators were designed and analyzed numerically for a single-axis prototype linear stage driven by a coreless linear motor. A magnetic circuit model was derived for the magnetic actuators. The static stiffness and load capacity of the air bearing stage in the vertical (magnetically preloaded) direction were experimentally measured and compared with the results from the numerical analysis. Motion control laws for three degrees of freedom (i.e., vertical, pitch, and roll motions) were obtained with a high linearity and independence for each axis. The active compensation of three motion errors, the vertical, pitch and roll motion errors were performed through curve-fitting the three errors measured with combination of capacitive gap sensors and a laser interferometer. The errors were reduced from 1.09 to 0.11 μm for the vertical straightness error, from 9.42 to 0.18 arcsec for the pitch motion, and from 2.42 to 0.18 arcsec for the roll motion as level of measured repeatability.