Analysis of changes in the viability and gene expression profiles of human mesenchymal stromal cells over time.

BACKGROUND AIMS: Because of their ability to differentiate and widespread availability, human mesenchymal stromal cells (hMSC) are often used as a clinical therapeutic tool. However, the factors that determine the quality and viability of hMSC are not well understood. METHODS: We evaluated the viability of hMSC over time using flow cytometry analysis (FACS) and transmission electron microscopy (TEM) to determine if morphologic changes occurred in hMSC. In addition, we conducted gene expression prof ling using an Affymetrix Human Genome U133 Plus 2.0 Array. RESULTS: FACS analysis revealed that 83% and 76% of the cells were viable in sterilized phosphate-buffered saline (PBS) after 6 h and 12 h, respectively. TEM data revealed that the total number of cells with healthy chromatins or a few cytosolic vacuoles was significantly reduced over time. We then conducted gene expression profiling using a microarray, which revealed changes in the expression of 2949 functional genes. Specifically, among the total of 50,000 gene probes evaluated, the expression levels of apoptosis and stress-related genes were significantly increased over time. CONCLUSIONS: The results of this study suggest that the viability of hMSC decreases after disassociation from the culture dish and time is an essential factor when considering hMSC as a potential source for stem cell-based direct transplantation.

Anti-oxidative stress effect of red ginseng in the brain is mediated by peptidyl arginine deiminase type IV (PADI4) repression via estrogen receptor (ER) ß up-regulation.

AIM OF THE STUDY: Ginseng has been used as an anti-stress agent, and its active ingredient, ginsenoside, is similar in structure to estrogen. However, the effect of ginseng on the stressed brain is not completely understood. The aim of this study is to understand systematically how red ginseng (RG) affects gene expressions in the brain of immobilization (IMO) stressed mice to elucidate its underlying mechanism. MATERIALS AND METHODS: For in vivo experiments, mice were stressed by immobilization for 30, 45, or 60 min, and gene expression in the mice brain was analyzed by microarray and system biology. Apoptosis was measured by terminal deoxynucleotidyl transferase-mediated digoxigenin-dUTP nick-end labeling (TUNEL) staining, and gene expression by Western blot or qPCR. For in vitro study, the SK-N-SH neuroblastoma cells were stressed by H2O2 exposure. The resultant cytotoxicity was measured by MTT assay, and gene expression by Western blot, ELISA, or qPCR. RESULTS: Microarray analysis of genes in IMO stressed mice brains showed that RG administration prior to IMO stress downregulated >40 genes including peptidyl arginine deiminase type 4 (PADI4). Interestingly, PADI4 was up-regulated by various stresses such as H2O2, acrylamide, and tunicamycin in neuroblastoma SK-N-SH cells but inhibited by RG. IMO stress and in vitro H2O2 stress depressed the estrogen receptor (ER)-ß expression but not ERα. However, RG treatment increased ERß expression both in vivo and in vitro. Comparative analysis regarding the networks by systems biology revealed that TNF-α plays a critical role in IMO stress, and the cell death associated network was much higher than other categories. Consistently, the IMO stress induced TNF-α and Cox-2 expressions, malondialdehyde (MDA), and cell death in the brain, whereas RG administration inhibited these inductions in vivo. siRNA and transient expression studies revealed that ERß inhibited the PADI4 expression. CONCLUSION: PADI4 could be used as an oxidative stress marker. RG seems to inhibit oxidative stress-inducible PADI4 by up-regulating ERß expression in the brain thus protecting brain cells from apoptosis.

Korean Red Ginseng Up-regulates C21-Steroid Hormone Metabolism via Cyp11a1 Gene in Senescent Rat Testes.

Ginseng (Panax ginseng Meyer) has been shown to have anti-aging effects in animal and clinical studies. However, the molecular mechanisms by which ginseng exerts these effects remain unknown. Here, the anti-aging effect of Korean red ginseng (KRG) in rat testes was examined by system biology analysis. KRG water extract prepared in feed pellets was administered orally into 12 month old rats for 4 months, and gene expression in testes was determined by microarray analysis. Microarray analysis identified 33 genes that significantly changed. Compared to the 2 month old young rats, 13 genes (Rps9, Cyp11a1, RT1-A2, LOC365778, Sv2b, RGD1565959, RGD1304748, etc.) were up-regulated and 20 genes (RT1-Db1, Cldn5, Svs5, Degs1, Vdac3, Hbb, LOC684355, Svs5, Tmem97, Orai1, Insl3, LOC497959, etc.) were down-regulated by KRG in the older rats. Ingenuity Pathway Analysis of untreated aged rats versus aged rats treated with KRG showed that the affected most was Cyp11a1, responsible for C21-steroid hormone metabolism, and the top molecular and cellular functions are organ morphology and reproductive system development and function. When genes in young rat were compared with those in the aged rat, sperm capacitation related genes were down-regulated in the old rat. However, when genes in the old rat were compared with those in the old rat treated with KRG, KRG treatment up-regulated C21-steroid hormone metabolism. Taken together, Cyp11a1 expression is decreased in the aged rat, however, it is up-regulated by KRG suggesting that KRG seems enhance testes function via Cyp11a1.

Review of recycling performance indicators: a study on collection rate in Taiwan.

The Taiwan Environmental Protection Administration (Taiwan EPA) launched a national Extended Producer Responsibility (EPR) system after integrating eight private recycling organizations in 1998. After that, the environmental performance of the EPR system brought a lot of attention to policy makers. Many studies show positive environmental effects of the EPR system in Taiwan. However, there are controversial questions remained, such as whether the performance indicators used are the right choice to estimate the environmental effects of the recycling policy? Can those estimated results really reflect the performance of the system? This paper would therefore like to more accurately evaluate the performance indicators of the EPR system based on data observed over the past decade in Taiwan. In the process of evaluating the performance indicators, we have found that the collection rates for durable goods are often ignored in countries that pursue a zero waste policy. This may affect the actual recycling outcome and resource direction targeted by producers. However, in order for the collection rate to be adopted as a policy indicator, how to estimate the amounts of retired or waste products during a period is critical. In this paper, we estimate the collection rate for electrical and electronic waste by using the survival analysis and ownership data analysis approaches. We also provide a comparison of both approaches and put forward suggestions for directions in the future in solid waste management.