Evaluation of drug toxicity with hepatocytes cultured in a micro-space cell culture system

A micro-space cell culture system was recently developed in which cells such as hepatocytes can be cultured and formed into a multicellular three-dimensional (3D) architecture. In this study, we assessed the performance of HepG2 cells cultured in this micro-space cell culture system in a drug toxicity test, and evaluated the effects of micro-space culture on their hepatocyte-specific functions. The micro-space cell culture facilitated the formation of 3D HepG2 cell architecture. HepG2 cells cultured in a micro-space culture plate exhibited increased albumin secretion and enhanced mRNA expression levels of cytochrome P450 (CYP) enzyme compared to those cultured in a monolayer culture. When the cells were exposed to acetaminophen, a hepatotoxic drug, the damage to the HepG2 cells grown in micro-space culture was greater than the damage to the HepG2 cells grown in monolayer culture. In addition, human primary hepatocytes grown in micro-space culture also exhibited increased albumin secretion, enhanced CYP mRNA expression levels and increased sensitivity to acetaminophen compared to those grown in monolayer culture. These results suggest that this micro-space culture method enhances the hepatocyte-specific functions of hepatocytes, including drug-metabolizing enzyme activities, making hepatocytes grown in the micro-space culture system a useful tool for evaluating drug toxicity in vitro.     

Investigation of the Aroma Profile of Green Tea Leaves Using Organic Synthesis and Conventional Analytical Techniques

Although many aroma components have been identified in green tea leaves, the aroma compounds contributing to green tea＇s characteristic odor have not yet been reported. The authors recently reported that aroma components with a matcha-like odor are present in both green tea and black tea prepared from the Sayamakaori tea cuttivar. This matcha-like odor is similar to the odor of commercial available matcha （high-quality powdered green tea）, and is a specific odor feature of green tea leaves. At present, the green-tea odor is thought to arise from the combination of a large number of constituents. Recent reports indicate that a complex interaction between olfactory receptors and odorants is important for the evaluation of the odors. Taking into consideration these findings, the authors investigated the aroma profile of green tea, focusing on the characteristic molecular structures of the constituents that give matcha-like odor. Using a combination of organic synthesis and gas chromatography-mass spectrometry plus gas chromatography-olfactometry, the authors identified aroma components with matcha-like odors in five other tea cultivars. This investigation also revealed that several compounds with a formyl group were important constituents of the aroma of green tea leaves, although the odor of each constituent was not individually similar to the tea＇s overall aroma. The authors found for the first time a group of key components that have the matcha-like odor.     

Metastatic Voyage of Ovarian Cancer Cells in Ascites with the Assistance of Various Cellular Components

Epithelial ovarian cancer (EOC) is the most lethal gynecologic malignancy and has a unique metastatic route using ascites, known as the transcoelomic root. However, studies on ascites and contained cellular components have not yet been sufficiently clarified. In this review, we focus on the significance of accumulating ascites, contained EOC cells in the form of spheroids, and interaction with non-malignant host cells. To become resistant against anoikis, EOC cells form spheroids in ascites, where epithelial-to-mesenchymal transition stimulated by transforming growth factor-β can be a key pathway. As spheroids form, EOC cells are also gaining the ability to attach and invade the peritoneum to induce intraperitoneal metastasis, as well as resistance to conventional chemotherapy. Recently, accumulating evidence suggests that EOC spheroids in ascites are composed of not only cancer cells, but also non-malignant cells existing with higher abundance than EOC cells in ascites, including macrophages, mesothelial cells, and lymphocytes. Moreover, hetero-cellular spheroids are demonstrated to form more aggregated spheroids and have higher adhesion ability for the mesothelial layer. To improve the poor prognosis, we need to elucidate the mechanisms of spheroid formation and interactions with non-malignant cells in ascites that are a unique tumor microenvironment for EOC.     

Direct ethanol production from barley beta-glucan by sake yeast displaying Aspergillus oryzae beta-glucosidase and endoglucanase

Three beta-glucosidase- and two endoglucanase-encoding genes were cloned from Aspergillus oryzae, and their gene products were displayed on the cell surface of the sake yeast, Saccharomyces cerevisiae GRI-117-UK. GRI-117-UK/pUDB7 displaying beta-glucosidase AO090009000356 showed the highest activity against various substrates and efficiently produced ethanol from cellobiose. On the other hand, GRI-117-UK/pUDCB displaying endoglucanase AO090010000314 efficiently degraded barley beta-glucan to glucose and smaller cellooligosaccharides. GRI-117-UK/pUDB7CB codisplaying both beta-glucosidase AO090009000356 and endoglucanase AO090010000314 was constructed. When direct ethanol fermentation from 20 g/l barley beta-glucan as a model substrate was performed with the codisplaying strain, the ethanol concentration reached 7.94 g/l after 24 h of fermentation. The conversion ratio of ethanol from beta-glucan was 69.6% of the theoretical ethanol concentration produced from 20 g/l barley beta-glucan. These results showed that sake yeast displaying A. oryzae cellulolytic enzymes can be used to produce ethanol from cellulosic materials. Our constructs have higher ethanol production potential than the laboratory constructs previously reported.     

Vitamin E reduces cholesterol esterification and uptake of acetylated low density lipoprotein in macrophages

The effects of vitamin E on cholesteryl ester (CE) metabolism in 1774 cells were examined. Pretreatment of 1774 cells with vitamin E at concentrations above 50 μM significantly decreased acetylated low density lipoprotein (LDL)-induced incorporation of [¹⁴C]oleate into CF in cells in a dose-dependent manner. This was partly due to vitamin E Also significantly inhibiting the uptake of [³H]CE-labeled acetylated LDL by 1774 cells. A trend existed toward suppression of acyl-CoA:cholesterol acyltransferase (ACAT) activity in the cell lysate at high vitamin E concentration, but there was no effect on hydrolysis of CE. These data indicate that vitamin E reduces the uptake of modified LDL and suppresses ACAT activity, resulting in less cholesterol esterification in macrophages; a novel mechanism underlying the antiatherogenic properties of vitamin E.     

Synthesis and Photocrosslinking Reaction of N-Allylcarbamoylmethyl Cellulose Leading to Hydrogel

Summary The reaction of the carboxymethyl cellulose sodium salt (Na-CMC) (degrees of substitution (DS) = 1.2) with N-hydroxysuccinimide (Su-OH) in the presence of 1-hydroxybenzotriazole and 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide (EDC) was carried out in water to obtain the Su-OH ester of carboxymethyl cellulose, Su-CMC, with the DS values of 0.19 – 1.04. N-Allylcarbamoylmethyl cellulose (Allyl-CMC), which was prepared from the reaction of Su-CMC with an excess amount of allylamine, was crosslinked by UV-irradiation. In addition, the photocrosslinked Allyl-CMC film was swollen with water to form a hydrogel having a relatively high water-swelling property, e.g., the degree of swelling (ds) was ca. 360% for Allyl-CMC with the DS of 0.93.     

Enhanced production of hydroxymethylfurfural from fructose with solid acid catalysts by simple water removal methods

This paper demonstrates two simple ways to increase 5-hydroxymethylfurfural (HMF) yield (selectivity) in fructose dehydration with various solid acid catalysts. One is a water removal from the reaction mixture by a mild evacuation at 0.97 × 10⁵ Pa; it increases HMF yield for various catalysts (heteropoly acid, zeolite, and acidic resin). The removal of water suppresses two undesired reactions: the hydrolysis of HMF to levulinic acid and the reaction of partially dehydrated intermediates to condensation products. The other method is a decrease in the particle (bead) size of the resin (Amberlyst-15). The crushed and sieved Amberlyst-15 powder in a size of 0.15–0.053 mm shows 100% HMF yield at high fructose concentration (50 wt.% in DMSO), which is to our knowledge the highest yield to date. Near-infrared spectroscopic characterization of adsorbed water suggests that the enhanced yield can be caused by an improved removal of adsorbed water in a small-size resin particle.     

A new control methodology of wind turbine generators for frequency control of power system in isolated island

A wind turbine generator (WTG) system's output is not constant and fluctuates depending on wind conditions. Fluctuating power causes frequency deviations and adverse effects to an isolated power system when large output power from WTG systems is penetrated in the power system. This paper presents an output power control methodology of a WTG for frequency control using a load power estimator. The load power is estimated by a disturbance observer, and the output power command of the WTG is determined according to the estimated load. Besides, the WTG can also be controlled during wind turbulence since the output power command is determined by considering wind conditions. The reduction of the power system frequency deviation by using the WTG can be achieved by the proposed method. The effectiveness of the proposed method is validated by numerical simulations. Copyright © 2010 John Wiley & Sons, Ltd.     

Output power dispatch control for a wind farm in a small power system

Nowadays, a wind turbine generator (WTG) is required to provide control capabilities as the output power of WTG fluctuates. Under this scenario, this paper proposes an output power control method of a wind farm (WF) connected to a small power system using pitch angle control. In this control approach, the WF output power control is achieved by two control levels: central and local. In the central control, the WF output power command is determined by considering the frequency deviations and wind speeds using a fuzzy function. Then, the local output power commands for each of the WTGs are based on the proposed dispatch control. In the proposed dispatch control, the output commands of each WTG are determined by considering wind conditions for each of the WTGs. The simulation results by using an actual detailed model for the wind power system show the effectiveness of the proposed method. Copyright © 2010 John Wiley & Sons, Ltd.