Epigenetic control of the expression of a primate-specific microRNA cluster in human cancer cells.

Many of the known microRNAs (miRNAs) are encoded by polycistronic transcripts and are thought to be co-expressed. In this study, we discovered that the expression of a large miRNA cluster (C19MC) on human chromosome 19 is upregulated by the demethylating agent, 5-aza-2'-deoxycytidine (5-Aza-dC), in AGS gastric cancer cells. We found that C19MC was rarely expressed in most cells, but its expression was restored through DNA demethylation. We confirmed that this miRNA cluster was mainly expressed in the placenta, as previously reported. Furthermore, its expression pattern was highly correlated with the methylation state of a distal CpG-rich region located about 17.6 kb upstream of the miRNA cluster. Using combined bisulfite restriction analysis (COBRA) and bisulfite-sequencing techniques, we determined that this CpG-rich region is hypermethylated in the AGS and HeLa cells, but hypomethylated in the placenta tissue. In conclusion, we demonstrated that the expression pattern of the C19MC was activated in human cancer cells through demethylation of a CpG-rich region.

Interfacial energy levels and related properties of atomic-layer-deposited Al2O3 films on nanoporous TiO2 electrodes of dye-sensitized solar cells.

Low-temperature (approximately 150 degrees C), atomic-layer-deposited Al(2)O(3) films on nanoporous TiO2 electrodes of dye-sensitized solar cells (DSSCs) were investigated using electron spectroscopy. The power conversion efficiency (PCE) of the DSSCs was increased from 5.7% to 6.5%, an improvement of 14%, with one monolayer of Al(2)O(3) with a thickness of approximately 0.2 nm. The formation of Ti-O-Al(OH)(2) and interfacial dipole layers exhibited a strong influence on the work function of the Al(2)O(3) over-layers, while the thicker Al(2)O(3) over-layers caused the values of valence band maximum and band gap to approach the values associated with pure Al(2)O(3). A work function difference (Delta Phi(A-T)) of 0.4 eV and a recombination barrier height (epsilon(RB)) of 0.1 eV were associated with the highest PCE achieved by the first monolayer of the Al(2)O(3) layer. Thicker Al(2)O(3) over-layers, however, caused significant reduction of PCE with negative Delta Phi(T-A) and increased interfacial energy barrier height ((*)epsilon(IB)) between the N719 dyes and TiO2 electrodes. It was concluded that the PCE of the DSSCs may correlate with Delta Phi(A-T), epsilon(RB), and (*)epsilon(IB) resulting from various thicknesses of the Al(2)O(3) over-layers and that interfacial reactions, such as the formation of Ti-O-Al(OH)(2) and dipole layers, play an important role in determining the interfacial energy levels required to achieve optimal performance of dye-sensitized TiO2 solar cells.

Diabetes reduces aortic endothelial gap junctions in ApoE-deficient mice: simvastatin exacerbates the reduction.

We examined the endothelial gap junctions in diabetic hyperlipidemic mice. Male apolipoprotein E (apoE)-deficient mice were made diabetic by streptozotocin. Three weeks later, the animals were treated with simvastatin for 2 weeks. The expression of aortic gap junctions in the non-diabetic (n=10), untreated diabetic (n=10), and simvastatin-treated diabetic animals (n=6) was analyzed. There was a >4-fold increase in serum cholesterol level and >50% increase in plaque areas in the diabetic mice, regardless of simvastatin treatment. Western blotting of aortae showed reduced expression of connexin37 (Cx37) and Cx40 in the diabetic mice, which were further decreased in the simvastatin-treated diabetic mice. Immunoconfocal microscopy showed that endothelial gap junctions made of Cx37 and Cx40 were both reduced in the untreated diabetic mice compared with the non-diabetic mice (decrease: Cx37, 41%; Cx40, 42%; both p<0.01). The reduction was greater in the simvastatin-treated mice (decrease in treated diabetic vs non-diabetic: Cx37, 61%; Cx40, 79%; both p<0.01; decrease in treated diabetic vs untreated diabetic: Cx37, 34%; Cx40, 63%; both p<0.01). Cx37 and Cx40 were decreased in the endothelium of plaque surface. Cx43 appeared in the medial layer and inner layer of the intima. All three connexins were rarely expressed in monocytes/macrophages inside the plaques. In conclusion, in apoE-deficient mice, streptozotocin-induced diabetes is associated with downregulation of endothelial Cx37 and Cx40 gap junctions. Short-term treatment with simvastatin exacerbates the downregulation.

Identification of collapsin response mediator protein-2 as a potential marker of colorectal carcinoma by comparative analysis of cancer cell secretomes.

The cancer cell secretome may contain many potentially useful biomarkers. We therefore sought to identify proteins in the conditioned media of colorectal carcinoma (CRC) cell lines but not in those from other cancer cell lines. The secretomes of 21 cancer cell lines derived from 12 cancer types were analyzed by SDS-PAGE combined with MALDI-TOF MS. Among the 325 proteins identified, collapsin response mediator protein-2 (CRMP-2) was chosen for evaluation as a potential CRC biomarker, since it was selectively detected in the CRC cell line secretome and has never been reported as a cancer biomarker. Immunohistochemical analysis of 169 CRC specimens showed that CRMP-2 was positively detected in 58.6% of the tumors, but weakly or not detected in >90% of the adjacent nontumor epithelial cells. Moreover, the CRMP-2-positive rate was significantly increased in earlier stage tumors and lymph node metastasis. Plasma CRMP-2 levels were significantly higher in CRC patients (N = 201) versus healthy controls (N = 201) (61.3 +/- 34.6 vs. 40.2 +/- 24.3 ng/mL, p = 0.001). Our results indicate that comparative analysis of cancer cell secretome is a feasible strategy for identifying potential cancer biomarkers, and that CRMP-2 may be a novel CRC biomarker.

Monitoring the hygene of chicken hatcheries in Taiwan during 1999-2001.

Microorganism contamination in hatcheries and eggs has a serious impact on the viability and quality of chicks as well as on the overall growth performance of chickens. Microbiological agents are present in the fluff when chicks hatch. Detecting microorganisms in fluff is a convenient method for evaluating the hygienic status in a hatchery. Fluff samples from 31 hatcheries collected over 3 years were tested for the total bacterial count, the presence of Salmonella spp., and fungus to evaluate the hygienic status of hatcheries in Taiwan from 1999 through 2001. The total bacterial score from the fluff samples was calculated and expressed as a bacterial score in a log scale. Most hatcheries had a bacterial count ranged from scale 1 to 3. Among the hatcheries, 13% to 29% were contaminated with Salmonella spp.; and 33% to 73% were contaminated with fungi in different quarters. The third quarter of each year was the most contaminated period (p<0.01). According to the data obtained from the fluff tests, hatcheries keep their hygienic status and supply good quality chicks by cleaning and disinfecting.