Effect of surface properties of silica nanoparticles on their cytotoxicity and cellular distribution in murine macrophages

Surface properties are often hypothesized to be important factors in the development of safer forms of nanomaterials (NMs). However, the results obtained from studying the cellular responses to NMs are often contradictory. Hence, the aim of this study was to investigate the relationship between the surface properties of silica nanoparticles and their cytotoxicity against a murine macrophage cell line (RAW264.7). The surface of the silica nanoparticles was either unmodified (nSP70) or modified with amine (nSP70-N) or carboxyl groups (nSP70-C). First, the properties of the silica nanoparticles were characterized. RAW264.7 cells were then exposed to nSP70, nSP70-N, or nSP70-C, and any cytotoxic effects were monitored by analyzing DNA synthesis. The results of this study show that nSP70-N and nSP70-C have a smaller effect on DNA synthesis activity by comparison to unmodified nSP70. Analysis of the intracellular localization of the silica nanoparticles revealed that nSP70 had penetrated into the nucleus, whereas nSP70-N and nSP70-C showed no nuclear localization. These results suggest that intracellular localization is a critical factor underlying the cytotoxicity of these silica nanoparticles. Thus, the surface properties of silica nanoparticles play an important role in determining their safety. Our results suggest that optimization of the surface characteristics of silica nanoparticles will contribute to the development of safer forms of NMs.     

Reaction of d-glucose in water at high temperatures (410 °C) and pressures (180 MPa) for the production of dyes and nano-particles

An autoclave (120-mL) and an optical micro-reactor (50-nL) were used to study the hydrothermal decomposition of d-glucose at high temperatures and high pressures. During slow heating (0.18 °C/s) to 350 °C in the autoclave, water-soluble glucose (0.9 M) began to decompose at 220 °C and reacted completely at 280 °C. The initial decomposition products were 5-(hydroxymethyl)furfural and levoglucosan, and these subsequently converted into oil and solid residue, and finally to solid particles at a 65 wt% yield at 350 °C. When the same heating rate and temperature were used on glucose solutions in the micro-reactor, yellow and orange materials decomposed from glucose were produced. Numerous particles precipitated at 251 °C, and at 350 °C, all the glucose changed to an orange film and solid particles, which were nanoparticles as confirmed by SEM. However, when the glucose solution was rapidly heated to 410 °C (9.5-17 °C/s), yellow, brown and orange sugar-like materials were produced. A homogeneous phase with yellow color still remained at temperatures as high as 380 °C, and few particles formed until 410 °C. It can be concluded that micron-sized particles and colored solutions can be produced by slow heating, while rapid heating resulted in the formation of dye-like substances with glucose-like structures. The formation of colored solutions and particles may have technological implications in food or materials formation processes that use high temperature water with biomass feedstocks.     

Presence of Apolipoprotein C-III Attenuates Apolipoprotein E-Mediated Cellular Uptake of Cholesterol-Containing Lipid Particles by HepG2 Cells

Apolipoprotein C-III (apoC-III) decreases the apolipoprotein E (apoE)-mediated uptake of lipoprotein remnants by the liver, and a high plasma concentration of apoC-III in VLDL is associated with hypertriglyceridemia and the risk of coronary heart disease. In this study, we prepared lipid emulsions containing triolein, phosphatidylcholine and cholesterol as model particles of lipoproteins, and examined the roles of apoC-III in apoE-mediated uptake of emulsions by HepG2 cells. Cholesterol in emulsion particles enhanced the apoE-mediated uptake via heparan sulfate proteoglycan and LDL receptor-related protein pathways. The amount of apoE bound to emulsion particles was increased by the presence of cholesterol at the particle surface, whereas cholesterol had no effect on the binding amount of apoC-III. Surface cholesterol alleviated the inhibitory effect of apoC-III on apoE incorporation into the emulsion surface. However, ApoC-III almost completely inhibited the apoE-mediated uptake of cholesterol-containing emulsions despite sufficient binding of apoE to emulsions. These findings suggest that apoC-III attenuates the binding of apoE to the lipoprotein surface and apoE-mediated cellular uptake of lipoprotein remnants. Furthermore, cholesterol may affect these functions of apoC-III and apoE involved in the clearance of lipoprotein remnants.     

High-level production of prourokinase-annexin V chimeras in the methylotrophic yeast Pichia pastoris

Prourokinase (proUK)-annexin V chimeras expressed by the methylotrophic yeast Pichia pastoris in a synthetic medium as part of a system designed to yield a novel thrombolytic agent are degraded, as it is thought, by various yeast proteases present in the culture supernatant. Minimization of proteolysis was therefore investigated to increase the yield of intact proUK-annexin V. Protease inhibitor screening study indicated several proteases including at least serine protease like chymotrypsin were involved in the proteolysis. Addition of more than 10% of peptone or more than 0.2 mol l(-1) of arginine to the medium was effective in minimizing proteolysis in shake-flask culture. Culture condition of higher pH was also effective, however, induced a cell death. Cell improvement by increasing the methanol utilization ability yielded greater tolerance to high pH. As a result, the culture condition with highly concentrated peptone solution fed under controlled conditions of pH 8.0 was established, which greatly reduced proteolytic degradation in fed-batch fermentation. These optimal conditions, which enabled fibrinolytic activity to reach 7800 IU ml(-1), could easily be applied in industrial scale production.     

Dough properties and breadmaking qualities of whole waxy wheat flour and effects of additional enzymes

BACKGROUND: Waxy wheat, a new kind of genetically back‐crossed wheat, was applied to make whole bread in this study. Dough properties and bread quality of the whole waxy wheat flour, which was milled from 100% whole grains containing bran and germ, were determined. RESULTS: Whole waxy wheat had lower protein and lipid contents but higher dietary fiber content than whole regular wheat flour. Pasting temperature and viscosity of the whole waxy wheat flour were significantly lower than those of the whole regular wheat. However, the white wheat flour milled from wheat grains with 48% recovery had significantly higher peak viscosity than the whole waxy wheat. Bread made from the whole waxy wheat flour was significantly softer than that from the whole regular wheat flour during storage. However, bread made from whole waxy wheat had significantly lower specific volume than that from the white waxy flour because of the high amount of dietary fiber. Addition of cellulase increased paste viscosity, lowered dough mixing properties and reduced the firmness of the bread. The addition of pentosanase also increased paste viscosity, lowered dough mixing properties, improved loaf volume of bread but increased the firmness of breadcrumbs, while the addition of α‐amylase only increased final viscosity of flour and did not affect dough properties and bread qualities of whole waxy wheat flour. CONCLUSION: As a result, waxy wheat shows superior properties for making whole breads. Additional enzymes are also necessary to improve bread quality and nutritive values of whole waxy bread. Copyright © 2007 Society of Chemical Industry     

Contamination levels and congener distributions of PCDDs, PCDFs and Co-PCBs in several fast foods in Japan

We determined the concentrations of polychlorinated dibenzo-p-dioxins (PCDDs), dibenzofurans (PCDFs) and coplanar polychlorinated biphenyls (Co-PCBs) in three types of fast foods [(1) seventeen hamburgers and two hot dogs, (2) six portions of fried potatoes and (3) three chicken products] obtained from fast food shops or convenience stores in Japan. All samples tested showed low toxic equivalent quantity (TEQ) levels of dioxins in the range of 0.001-0.083 pg-TEQ/g wet weight (0.006-0.053 pg-TEQ/g for hamburgers and hot dogs, 0.001-0.083 pg-TEQ/g for fried potatoes and 0.053-0.065 pg-TEQ/g for chicken products). The congener profile in hamburgers and hot dogs suggested that the total TEQ was mainly determined by Co-PCBs, especially by 3,3',4,4',5-PeCB (#126), which accounted for 44% of the total TEQ value. Findings for animal foods such as beef and cheese were consistent with this result. For fried potatoes, PCDD/Fs accounted for 94% of the total TEQ value, and 2,3,4,7,8-PeCDF accounted for 32% of PCDD/Fs. Dioxins in the chicken products consisted of 3,3',4,4',5-PeCB (#126) and 1,2,3,7,8-PeCDD, which accounted for 23% and 21% of the total TEQ, respectively. If an adult (50 kg weight) eats 150 g of hamburger, 100 g of potatoes and 150 g of chicken, the daily intake is estimated to be 0.299 pg-TEQ/kg b.w./day using the average values (0.022, 0.028 and 0.059 pg-TEQ/g, respectively) obtained in this study. This value corresponds to 7.5% of the tolerable daily intake (TDI) for PCDD/Fs and Co-PCBs in Japan.     

Hydrogen and ethanol production from glycerol-containing wastes discharged after biodiesel manufacturing process

H2 and ethanol production from glycerol-containing wastes discharged after a manufacturing process for biodiesel fuel (biodiesel wastes) using Enterobacter aerogenes HU-101 was evaluated. The biodiesel wastes should be diluted with a synthetic medium to increase the rate of glycerol utilization and the addition of yeast extract and tryptone to the synthetic medium accelerated the production of H2 and ethanol. The yields of H2 and ethanol decreased with an increase in the concentrations of biodiesel wastes and commercially available glycerol (pure glycerol). Furthermore, the rates of H2 and ethanol production from biodiesel wastes were much lower than those at the same concentration of pure glycerol, partially due to a high salt content in the wastes. In continuous culture with a packed-bed reactor using self-immobilized cells, the maximum rate of H2 production from pure glycerol was 80 mmol/l/h yielding ethanol at 0.8 mol/mol-glycerol, while that from biodiesel wastes was only 30 mmol/l/h. However, using porous ceramics as a support material to fix cells in the reactor, the maximum H2 production rate from biodiesel wastes reached 63 mmol/l/h obtaining an ethanol yield of 0.85 mol/mol-glycerol.     

Ferulic acid production from clove oil by Pseudomonas fluorescens E118

For the production of the useful antioxidant ferulic acid from clove oil containing abundant eugenol, the growth of eugenol-degrading microorganisms in the presence of clove oil was examined. Pseudomonas fluorescens E118, a clove-oil-tolerant strain, accumulated 6.1 g/l ferulic acid under optimized culture conditions with the intermittent addition of eugenol. When the bacterium was applied to ferulic acid production from clove oil, 5.8 g/l ferulic acid was produced with the intermittent addition of clove oil. Since clove oil is much cheaper than eugenol, ferulic acid production from clove oil by the bacterium is promising for the industrial production of ferulic acid.     

Sensitivity of Diesel Particulate Material Emissions and Composition to Blends of Petroleum Diesel and Biodiesel Fuel

A number of investigations have examined the impact of the use of biodiesel on the emissions of carbon dioxide and regulated emissions, but limited information exists on the chemical composition of particulate matter from diesel engines burning biodiesel blends. This study examines the composition of diesel particulate matter (DPM) emissions from a commercial agriculture tractor burning a range of biodiesel blends operating under a load that is controlled by a power take off (PTO) dynamometer. Ultra-low sulfur diesel (ULSD) fuel was blended with soybean and beef tallow based biodiesel to examine fuels containing 0% (B0), 25% (B25), 50% (B50), 75% (B75), and 100% (B100) biodiesel. Samples were then collected using a dilution source sampler to simulate atmospheric dilution. Diluted and aged exhaust was analyzed for particle mass and size distribution, PM_2.5 particle mass, PM_2.5 organic and elemental carbon, and speciated organic compounds. PM_2.5 mass emissions rates for the B25, B50, and B75 soybean oil biodiesel mixtures had 20%–30% lower emissions than the petroleum diesel, but B100 emissions were about 40% higher than the petroleum diesel. The trends in mass emission rates with the increasing biodiesel content can be explained by a significant decrease in elemental carbon (EC) emissions across all blending ranges and increasing organic carbon (OC) emissions with pure biodiesel. Beef tallow biodiesel blends showed similar trends. Nevertheless, it is important to note that the study measurements are based on low dilution rates and the OC emissions changes may be affected by ambient temperature and different dilution conditions spanning micro-environments and atmospheric conditions. The results show that the use of biodiesel fuel for economic or climate change mitigation purposes can lead to reductions in PM emissions and a co-benefit of EC emission reductions. Detailed speciation of the OC emissions were also examined and are presented to understand the sensitivity of OC emissions with respect to biodiesel fuel blends.

Copyright 2012 American Association for Aerosol Research