Process design of chondrocyte cultures with monolayer growth for cell expansion and subsequent three-dimensional growth for production of cultured cartilage

The subculture of rabbit chondrocytes with serial passaging was carried out for cell expansion on a collagen-coated surface, and the morphological transition of round-shaped cells to spindle-shaped ones was examined. The observation of cytoskeletal formation by staining F-actin and vinculin supported the view that the round-shaped cells were in the process of differentiation with immature stress fibers relating to less cellular polarity. The cellular morphology was estimated in terms of the distribution of roundness, R(C), during the subculturing on the collagen substrate. The frequency of the number of round-shaped cells, which was defined as the ratio of the number of cells with R(C) >0.9 against the total cell number, was correlated in a logarithmic formula with the number of population doublings during the subcultures. Kinetic models were adopted for the process design of the combined culture of chondrocytes with monolayer growth on the collagen substrate and subsequent three-dimensional growth in Atelocollagen gel, employing the boundary conditions based on the population balance between differentiated and dedifferentiated cells. The combined culture was performed successfully according to the process design scheduled as monolayer growth for 240 h and three-dimensional growth for 264 h, the number of seed cells being 68% of that in the conventional culture for 504 h where monolayer growth for cell expansion was not included.     

Reduction in carbon dioxide emission, and enhancement of cell yield by control of light intensity in photomixotrophic batch culture of Marchantia polymorpha

Photomixotrophic cultures of Marchantia polymorpha were examined under light irradiation using glucose as an organic carbon source. The activity of ribulose 1,5-bisphosphate carboxylase/oxygenase in the cells was found to be maximum at an absorbed light energy of E(c) = 8.7 x 10(2)W/kg and the respiration rate of the cells remained at a low level in the range of E(c) = 1.7 x 10(2) to 1.5 x 10(3) W/kg. Batch culture of M. polymorpha was carried out in a bioreactor while keeping the E(c) value at about 3.5 x 10(2) W/kg by regulating incident light intensity between 19 and 220 W/m2. During the culture conducted under controlled light, CO2 evolution from the reactor was effectively suppressed and the obtained cell yield was 0.88 kg dry cells/kg glucose, whereas the yield was 0.62 kg dry cells/kg glucose throughout the culture conducted at the constant incident light intensity of 50 W/m2.     

Prediction of the stress—strain curve of a short-fibre reinforced thermoplastic

Analytical work has been performed to predict the stress—strain curve of a carbon fibre reinforced polyamide 66. A typical stress—strain curve of this composite consists of three stages. Beyond the first linear stage the change of slope of the curve is attributed to the initiation and extension of cracks at fibre ends. Several theoretical models have been developed to take into account the development of cracks at various loading levels. Fibre volume-fraction, aspect ratio and orientation as well as fibre and matrix elastic properties and interface energies are the major parameters considered. The theoretical predictions are in close agreement with the experimental data.     

First-order phase transition with a moving boundary. II. Theory of ice formation

An exact solution of the parabolic partial differential equation with a moving boundary condition is found. Applications to the problem of ice formation are discussed.     

Introducing the GASP scale: A new measure of guilt and shame proneness.

Although scholars agree that moral emotions are critical for deterring unethical and antisocial behavior, there is disagreement about how 2 prototypical moral emotions—guilt and shame—should be defined, differentiated, and measured. We addressed these issues by developing a new assessment—the Guilt and Shame Proneness scale (GASP)—that measures individual differences in the propensity to experience guilt and shame across a range of personal transgressions. The GASP contains 2 guilt subscales that assess negative behavior-evaluations and repair action tendencies following private transgressions and 2 shame subscales that assess negative self-evaluations (NSEs) and withdrawal action tendencies following publically exposed transgressions. Both guilt subscales were highly correlated with one another and negatively correlated with unethical decision making. Although both shame subscales were associated with relatively poor psychological functioning (e.g., neuroticism, personal distress, low self-esteem), they were only weakly correlated with one another, and their relationships with unethical decision making diverged. Whereas shame–NSE constrained unethical decision making, shame–withdraw did not. Our findings suggest that differentiating the tendency to make NSEs following publically exposed transgressions from the tendency to hide or withdraw from public view is critically important for understanding and measuring dispositional shame proneness. The GASP's ability to distinguish these 2 classes of responses represents an important advantage of the scale over existing assessments. Although further validation research is required, the present studies are promising in that they suggest the GASP has the potential to be an important measurement tool for detecting individuals susceptible to corruption and unethical behavior. (PsycINFO Database Record (c) 2011 APA, all rights reserved)     

A Study on High Subsonic Airfoil Flows in Relatively High Reynolds Number by Using OpenFOAM

In the present study, numerical calculations of the flow-field around the airfoil model are performed by using the OpenFOAM in high subsonic flows. The airfoil model is NACA 64A010. The maximum thickness is 10 % of the chord length. The SonicFOAM and the RhoCentralFOAM are selected as the solver in high subsonic flows. The grid point is 158,000 and the Mach numbers are 0.277 and 0.569 respectively. The CFD data are compared with the experimental data performed by the cryogenic wind tunnel in the past. The results are as follows. The nu- merical results of the pressure coefficient distribution on the model surface calculated by the SonicFOAM solver showed good agreement with the experimental data measured by the cryogenic wind tunnel. And the data calcu- lated by the SonicFOAM have the capability for the quantitative comparison of the experimental data at low an- gle of attack.     

Physical function was related to mortality in patients with chronic kidney disease and dialysis

Previous studies have shown that exercise improves aerobic capacity, muscular functioning, cardiovascular function, walking capacity, and health‐related quality of life (QOL) in patients with chronic kidney disease (CKD) and dialysis. Recently, additional studies have shown that higher physical activity contributes to survival and decreased mortality as well as physical function and QOL in patients with CKD and dialysis. Herein, we review the evidence that physical function and physical activity play an important role in mortality for patients with CKD and dialysis. During November 2016, Medline and Web of Science databases were searched for published English medical reports (without a time limit) using the terms “CKD” or “dialysis” and “mortality” in conjunction with “exercise capacity,” “muscle strength,” “activities of daily living (ADL),” “physical activity,” and “exercise.” Numerous studies suggest that higher exercise capacity, muscle strength, ADL, and physical activity contribute to lower mortality in patients with CKD and dialysis. Physical function is associated with mortality in patients with CKD and dialysis. Increasing physical function may decrease the mortality rate of patients with CKD and dialysis. Physicians and medical staff should recognize the importance of physical function in CKD and dialysis. In addition, exercise is associated with reduced mortality among patients with CKD and dialysis.     

Comprehension of terminal differentiation and dedifferentiation of chondrocytes during passage cultures

A high density collagen type I coated substrate (CL substrate) was used to evaluate the chondrocyte phenotypes in passaged cultures. With increasing age of cell population (population doubling (PD)=0-14.5), the frequency of non-dividing spindle shaped cells without ALP activity increased, accompanied with an increase in gene expression of collagen type I, meaning the senescence of dedifferentiated cells. At the middle age of cell population (PD=5.1 and 6.6), the high frequency of polygonal shaped cells with ALP activity existed on the CL substrate together with up-regulated expressions of collagen types II and X, indicating the terminal differentiation of chondrocytes. When the chondrocytes passaged up to the middle age were embedded in collagen gel, the high frequency of single hypertrophic cells with collagen type II formation was recognized, which supports the thought that the high gene expression of collagen type II was attributed to terminal differentiation rather than redifferentiation. These results show that the CL substrate can draw out the potential of terminal differentiation in chondrocytes, which is unattainable by a polystyrene surface, and that the CL substrate can be a tool to evaluate cell quality in three-dimensional culture with the collagen gel.     

A close correlation in the expression patterns of Af-6 and Usp9x in Sertoli and granulosa cells of mouse testis and ovary

Usp9x, an X-linked deubiquitylating enzyme, is stage dependently expressed in the supporting cells (i.e. Sertoli cells and granulosa cells) and germ cells during mouse gametogenesis. Af-6, a cell junction protein, has been identified as a substrate of Usp9x, suggesting a possible association between Usp9x and Af-6 in spermatogenesis and oogenesis. In this study, we examined the expression pattern of Af-6 and Usp9x and their intracellular localization in testes and ovaries of mice treated with or without pregnant mare serum gonadotropin (PMSG), an FSH-like hormone. In both testes and ovaries, Af-6 expression was predominantly observed in supporting cells, as well as in steroidogenic cells, but not in any germ cells. In Sertoli cells, Af-6 was continuously expressed throughout postnatal and adult stages, where both Af-6 and Usp9x were enriched at the sites of Sertoli-Sertoli and Sertoli-spermatid junctions especially at stages XI-VI. In the granulosa cells, Af-6, as well as Usp9x, was highly expressed in primordial and primary follicles, but its expression rapidly decreased after the late-secondary follicle stage. Interestingly, in PMSG-treated mice, the expression levels of Af-6 and Usp9x were synchronously enhanced, slightly in Sertoli cells and strongly in granulosa cells of the late-secondary and Graafian follicles. Such closely correlated expression patterns between Af-6 and Usp9x clearly suggest that Af-6 may be deubiquitylated by Usp9x in both Sertoli and granulosa cells. It further suggests that the post-translational regulation of Af-6 by Usp9x may be one potential pathway to control the cell adhesion dynamics in mammalian gametogenesis.