Influence of production systems on phenolic characteristics and antioxidant capacity of highbush blueberry cultivars.

Blueberry is a crop grown worldwide due to the excellent quality and high polyphenol content of its fruit and tolerance to cold conditions. We investigated the influence of three production systems, namely an open field, heated greenhouse, and non-heated (plastic) greenhouse, on the phenolic characteristics (total phenolic, flavonoid, and anthocyanin content) and antioxidant capacities of "Spartan" (northern highbush), "Sharpblue" (southern highbush), and "O'Neal" (southern highbush) blueberry cultivars. The non-heated production system showed the highest phenolic characteristics and antioxidant capacity in "Spartan" and "O'Neal," while the open field production system showed the highest phenolic characteristics and antioxidant capacity in "Sharpblue." Derivatives of delphinidin and malvidin were two of the most abundant anthocyanins. The heated greenhouse production system resulted in larger amounts of delphinidin derivatives compared with the other production systems, while the blueberry grown in the non-heated greenhouse produced larger amount of malvidin derivatives. The anthocyanin profiles varied according to production system and blueberry cultivars. The principal component analysis loading plot of blueberries for individual anthocyanins explained over 95% of the total variance. In summary, the results of this study suggest that a strategic approach to blueberry production could elevate the phenolic content and antioxidant capacity of cultivated blueberry. PRACTICAL APPLICATION: The highbush blueberry, a rich source of bioactive polyphenols, is a popular fruit. The microclimate of the production system of highbush blueberries affects the concentrations of antioxidative phenolic compounds such as anthocyanins. Therefore, discovering and applying the appropriate method of production for each blueberry cultivar could facilitate production of high-quality blueberries rich in phenolic antioxidants.

Physiochemical properties, dietary fibers, and functional characterization of three yuzu cultivars at five harvesting times.

This research focused on physiochemical and nutritional properties and functional characterization of three cultivars of yuzu-Native, Tadanishiki yuzu, and Namhae1-during different seasons. According to the cultivar and harvest time, yuzu cultivars were analyzed for free sugar, dietary fiber, hesperidin, naringin, and flavonoid content as well as antioxidant and antihypertensive activity. During November, Namhae1 exhibited the highest fruit weight, °Brix/acidity ratio, and total dietary fiber content. Tadanishiki contained the highest fructose and sucrose levels, pectin and cellulose contents, and soluble dietary fiber. Tadanishiki also had the highest hesperidin content in October, while the naringin content and antioxidant activity were the greatest in November. Antihypertensive activity was also the strongest for Tadanishiki, which was picked in October and November. These results indicated that Tadanishiki in October or November was the best for consumption or favorable processing because of its excellent product quality and high levels of nutritional and functional compounds.

Phenolic Profiles of Hardy Kiwifruits and Their Neuroprotective Effects on PC-12 and SH-SY5Y Cells against Oxidative Stress.

Hardy kiwifruits (Actinidia arguta Planch.) have high amounts of antioxidants, including ascorbic acid (vitamin C) and phenolics. The anti-cholinesterase activity and neuroprotective effects of three different cultivars of hardy kiwifruits, cv. Mansu (A. arguta × A. deliciosa), cv. Haeyeon (A. arguta), and cv. Chiak (A. arguta), on PC-12 and SH-SY5Y cells were evaluated. Extraction of phenolics and vitamin C was carried out using 80% (v/v) aqueous ethanol and metaphosphoric acid assisted with homogenization, respectively. Hardy kiwifruit of cv. Mansu showed higher total phenolic, total flavonoid, and vitamin C contents and antioxidant capacity compared to the other tw°Cultivars of hardy kiwifruits, cv. Haeyeon and cv. Chiak. Analysis of high-performance liquid chromatography results revealed the presence of procyanidin B2, (?)-epicatechin, neochlorogenic acid, cryptochlorogenic acid, rutin, hyperoside, isoquercitrin, and astragalin in hardy kiwifruits. The three cultivars of hardy kiwifruits had a wide range of vitamin C content of 55.2?130.0 mg/100 g fresh weight. All three cultivars of hardy kiwifruits had protective effects on neuronal PC-12 and SHSY5Y cells exposed to hydrogen peroxide by increasing cell viability and reducing intracellular oxidative stress. Furthermore, the hardy kiwifruits inhibited acetylcholinesterase and butyrylcholinesterase. Collectively, these results suggest that hardy kiwifruits rich in antioxidants like phenolics and vitamin C have good potential as functional materials in neuroprotective applications.

Pentamidine Inhibits Titanium Particle-Induced Osteolysis In Vivo and Receptor Activator of Nuclear Factor-κB Ligand-Mediated Osteoclast Differentiation In Vitro.

Background: Wear debris-induced osteolysis leads to periprosthetic loosening and subsequent prosthetic failure. Since excessive osteoclast formation is closely implicated in periprosthetic osteolysis, identification of agents to suppress osteoclast formation and/or function is crucial for the treatment and prevention of wear particle-induced bone destruction. In this study, we examined the potential effect of pentamidine treatment on titanium (Ti) particle-induced osteolysis, and receptor activator of nuclear factor-κB ligand (RANKL)-induced osteoclastogenesis. Methods: The effect of pentamidine treatment on bone destruction was examined in Ti particle-induced osteolysis mouse model. Ti particles were implanted onto mouse calvaria, and vehicle or pentamidine was administered for 10 days. Then, calvarial bone tissue was analyzed using micro-computed tomography and histology. We performed in vitro osteoclastogenesis assay using bone marrow-derived macrophages (BMMs) to determine the effect of pentamidine on osteoclast formation. BMMs were treated with 20 ng/mL RANKL and 10 ng/mL macrophage colony-stimulating factor in the presence or absence of pentamidine. Osteoclast differentiation was determined by tartrate-resistant acid phosphatase staining, real-time polymerase chain reaction, and immunofluorescence staining. Results: Pentamidine administration decreased Ti particle-induced osteoclast formation significantly and prevented bone destruction compared to the Ti particle group in vivo. Pentamidine also suppressed RANKL-induced osteoclast differentiation and actin ring formation markedly, and inhibited the expression of nuclear factor of activated T cell c1 and osteoclast-specific genes in vitro. Additionally, pentamidine also attenuated RANKL-mediated phosphorylation of IκBα in BMMs. Conclusion: These results indicate that pentamidine is effective in inhibiting osteoclast formation and significantly attenuates wear debris-induced bone loss in mice.

Inhibitory effects of methyl-3,5-di-O-caffeoyl-epi-quinate on RANKL-induced osteoclast differentiation.

In this study, we have shown that methyl-3,5-di-O-caffeoyl-epi-quinate, a naturally occurring compound isolated from Ainsliaea acerifolia, inhibits receptor activator of nuclear factor-κB ligand (RANKL)-induced formation of multinucleated tartrate-resistant acid phosphatase (TRAP)-positive osteoclasts and the expression of osteoclast marker genes. Methyl-3,5-di-O-caffeoyl-epi-quinate also inhibited RANKL-induced activation of p38, Akt and extracellular signal-regulated kinase (ERK) as well as the expression of nuclear factor of activated T-cell (NFATc1), the key regulator of osteoclast differentiation. Negative regulators for osteoclast differentiation was upregulated by methyl-3,5-di-O-caffeoyl-epi-quinate. Collectively, our results suggested that methyl-3,5-di-O-caffeoyl-epi-quinate suppresses osteoclast differentiation via downregulation of RANK signaling pathways and NFATc1.

Diphlorethohydroxycarmalol from Ishige okamurae Suppresses Osteoclast Differentiation by Downregulating the NF-κB Signaling Pathway.

Marine algae possess a variety of beneficial effects on human health. In this study, we investigated whether diphlorethohydroxycarmalol (DPHC), isolated from Ishige okamurae, a brown alga, suppresses receptor activator of nuclear factor-κB ligand (RANKL)-induced osteoclast differentiation. DPHC significantly suppressed RANKL-induced osteoclast differentiation and macrophage-colony stimulating factor (M-CSF) expression in a dose-dependent manner. In addition, it significantly inhibited actin ring formation, the expression of osteoclast marker genes, such as tartrate-resistant acid phosphatase (TRAP), nuclear factor of activated T-cells cytoplasmic 1 (Nfatc1), cathepsin K (Ctsk), and dendritic cell-specific transmembrane protein (Dcstamp), and osteoclast-induced bone resorption. Analysis of the RANKL-mediated signaling pathway showed that the phosphorylation of both IκB and p65 was specifically inhibited by DPHC. These results suggest that DPHC substantially suppresses osteoclastogenesis by downregulating the RANK-NF-κB signaling pathway. Thus, it holds significant potential for the treatment of skeletal diseases associated with an enhanced osteoclast activity.

Analysis of Chain Branch of Polyolefins by a New Proton NMR Approach.

The crystallinity of polyethylene, which significantly affects the properties of the polymer, is quite sensitive to the concentration of its branches. Thus, it is necessary to estimate branch concentration with reasonable accuracy. Currently, (13)C NMR and gel permeation chromatography-Fourier transform infrared spectroscopy are widely-used analysis methods for the analysis of branch concentration. Despite several advantages, these methods sometimes have limitations. For instance, the preparation of samples for (13)C- NMR is tedious because high-concentration samples are required and the time for analysis is greater than 12 h. To more efficiently estimate the branch concentration of polyethylene, we developed a new high-field (1)H NMR method with an improved peak resolution by employing (1) homonuclear decoupling and (2) 2D heteronuclear correlation. The new method was observed to significantly reduce the experimental time to ∼ 30 min; furthermore, sample preparation was relatively simple because the method did not require high-concentration samples.

High-throughput method of dioxin analysis in aqueous samples using consecutive solid phase extraction steps with the new C18 Ultraflow™ pressurized liquid extraction and automated clean-up.

A high-throughput analytical method has been developed for the determination of seventeen 2,3,7,8-substituted congeners of polychlorinated dibenzo-p-dioxins and dibenzofurans (PCDD/Fs) in aqueous samples. A recently introduced octadecyl (C18) disk for semi-automated solid-phase extraction of PCDD/Fs in water samples with a high level of particulate material has been tested for the analysis of dioxins. A new type of C18 disk specially designed for the analysis of hexane extractable material (HEM), but never previously reported for use in PCDD/Fs analysis. This kind of disk allows a higher filtration flow, and therefore the time of analysis is reduced. The solid-phase extraction technique is used to change samples from liquid to solid, and therefore pressurized liquid extraction (PLE) can be used in the pre-treatment. In order to achieve efficient purification, extracts from the PLE are purified using an automated Power-prep system with disposable silica, alumina, and carbon columns. Quantitative analyses of PCDD/Fs were performed by GC-HRMS using multi-ion detection (MID) mode. The method was successfully applied to the analysis of water samples from the wastewater treatment system of a vinyl chloride monomer plant. The entire procedure is in agreement with EPA1613 recommendations regarding the blank control, MDLs (method detection limits), accuracy, and precision. The high-throughput method not only meets the requirements of international standards, but also shortens the required analysis time from 2 weeks to 3d.

Gas Chromatographic Method for the Analysis of Organic Acids in the Bio-Catalytic Conversion Process.

An analytical method for the quantification of acrylic acid (AA), 1,3-propanediol (1,3-PD) and 3-hydroxypropionic acid (3-HP) in the bio-catalytic conversion process has been developed by gas chromatography. A simple liquid-liquid extraction (LLE) procedure was used in the sample preparation. Organic acid additives such as trifluoroacetic acid were used to improve the extraction efficiency in the LLE procedure. Under optimum analysis conditions, all analytes were satisfactorily separated with no interference. In standard calibration, all correlation coefficients (r(2)) were better than or equal to 0.994. In culture media, the intra-batch precision (% relative standard deviation) and recovery (%) as the average value of the quality control samples were 2.3 and 102.4%, respectively. In addition, the inter-batch precision and recovery as the average value of the quality control samples were 5.0 and 104.0%, respectively. In phosphate buffer, the intra-batch precision and recovery as the average value of the quality control samples were 2.7 and 101.6%, respectively. In addition, the inter-batch precision and recovery as the average value of the quality control samples were 2.9 and 101.7%, respectively. The limit of detection (S/N ratio: 3) and limit of quantification (S/N ratio: 10) were 1.0 and 3.5 µg/mL, 3.0 and 10.0 µg/mL, and 9.0 and 30.0 µg/mL, respectively, for AA, 1,3-PD and 3-HP. Consequently, this method was demonstrated to be acceptable for the quantitative analysis of AA, 1,3-PD and 3-HP in culture media and phosphate buffer.

Effect of surface tension and coefficient of thermal expansion in 30 nm scale nanoimprinting with two flexible polymer molds.

We report on nanoimprinting of polymer thin films at 30 nm scale resolution using two types of ultraviolet (UV)-curable, flexible polymer molds: perfluoropolyether (PFPE) and polyurethane acrylate (PUA). It was found that the quality of nanopatterning at the 30 nm scale is largely determined by the combined effects of surface tension and the coefficient of thermal expansion of the polymer mold. In particular, the polar component of surface tension may play a critical role in clean release of the mold, as evidenced by much reduced delamination or broken structures for the less polarized PFPE mold when patterning a relatively hydrophilic PMMA film. In contrast, such problems were not notably observed with a relatively hydrophobic PS film for both polymer molds. In addition, the demolding characteristic was also influenced by the coefficient of thermal expansion so that no delamination or uniformity problems were observed when patterning a UV-curable polymer film at room temperature. These results suggest that a proper polymeric mold material needs to be chosen for patterning polymer films under different surface properties and processing conditions, providing insights into how a clean demolding characteristic can be obtained at 30 nm scale nanopatterning.

Shape-controllable microlens arrays via direct transfer of photocurable polymer droplets.

A simple method is presented to form an array of shape-controllable microlenses by partial photocuring of an UV-curable polymer and direct transfer. Using the transferred lens array, nanoscale metal patterns as small as 130-nm gaps are detected under an optical microscope with a distinguishable resolution.

High-throughput single-cell quantification using simple microwell-based cell docking and programmable time-course live-cell imaging.

Extracting single-cell information during cellular responses to external signals in a high-throughput manner is an essential step for quantitative single-cell analyses. Here, we have developed a simple yet robust microfluidic platform for measuring time-course single-cell response on a large scale. Our method combines a simple microwell-based cell docking process inside a patterned microfluidic channel, with programmable time-course live-cell imaging and software-aided fluorescent image processing. The budding yeast, Saccharomyces cerevisiae (S. cerevisiae), cells were individually captured in microwells by multiple sweeping processes, in which a cell-containing solution plug was actively migrating back and forth several times by a finger-pressure induced receding meniscus. To optimize cell docking efficiency while minimizing unnecessary flooding in subsequent steps, circular microwells of various channel dimensions (4-24 µm diameter, 8 µm depth) along with different densities of cell solution (1.5-6.0 × 10(9) cells per mL) were tested. It was found that the microwells of 8 µm diameter and 8 µm depth allowed for an optimal docking efficiency (>90%) without notable flooding issues. For quantitative single-cell analysis, time-course (time interval 15 minute, for 2 hours) fluorescent images of the cells stimulated by mating pheromone were captured using computerized fluorescence microscope and the captured images were processed using a commercially available image processing software. Here, real-time cellular responses of the mating MAPK pathway were monitored at various concentrations (1 nM-100 µM) of mating pheromone at single-cell resolution, revealing that individual cells in the population showed non-uniform signaling response kinetics.

Fluid-shear-stress-induced translocation of aquaporin-2 and reorganization of actin cytoskeleton in renal tubular epithelial cells.

In vivo, renal tubular epithelial cells are exposed to luminal fluid shear stress (FSS) and a transepithelial osmotic gradient. In this study, we used a simple collecting-duct-on-a-chip to investigate the role of an altered luminal microenvironment in the translocation of aquaporin-2 (AQP2) and the reorganization of actin cytoskeleton (F-actin) in primary cultured inner medullary collecting duct (IMCD) cells of rat kidney. Immunocytochemistry demonstrated that 3 h of exposure to luminal FSS at 1 dyn cm(-2) was sufficient to induce depolymerization of F-actin in those cells. We observed full actin depolymerization after 5 h exposure and substantial re-polymerization within 2 h of removing the luminal FSS, suggesting that the process is reversible and the fluidic environment regulates the reorganization of intracellular F-actin. We demonstrate that several factors (i.e., luminal FSS, hormonal stimulation, transepithelial osmotic gradient) collectively exert a profound effect on the AQP2 trafficking in the collecting ducts, which is associated with actin cytoskeletal reorganization.

Step-and-repeat process for thermal nanoimprint lithography.

We present a step-and-repeat process for thermal nanoimprint lithography. For the selective heating and imprinting, a spin-coated polystyrene layer is exposed to infra-red rays from a halogen lamp (intensity approximately 500 W) with a metal-covered glass while pressed with a transparent polymer mold (Young's modulus approximately 300 MPa) under a pressure of approximately 4 bar for 60-120 s. During imprinting, the non-irradiated region is protected by a metal screen and a heat sink consisting of a copper block at the bottom which prevents the pattern collapse by lateral heat conduction from the irradiated region.

Wetting transition and optimal design for microstructured surfaces with hydrophobic and hydrophilic materials.

We present wetting transition of a water droplet on microstructured polymer surfaces using materials with different hydrophilicity or hydrophobicity: hydrophobic polydimethyl siloxane (PDMS) (theta(water) approximately 110 degrees) and hydrophilic Norland Optical Adhesive (NOA) (theta(water) approximately 70 degrees). The microstructures were fabricated by replica molding and self-replication with varying pillar geometry [diameter: 5 microm, spacing-to-diameter ratio (s/d): 1-10 (equal interval), height-to-diameter ratio (h/d): 1-5] over an area of 100 mm(2) (10 mm x 10 mm). Measurements of contact angle (CA) and contact angle hysteresis (CAH) demonstrated that wetting state was either in the homogeneous Cassie regime or in the mixed regime of Cassie and Wenzel states depending on the values of s/d and h/d. These two ratios need to be adjusted to maintain stable superhydrophobic properties in the Cassie regime; s/d should be smaller than approximately 7 (PDMS) and approximately 6 (NOA) with h/d being larger than approximately 2 to avoid wetting transition by collapse of a water droplet into the microstructure. Based on our observations, optimal design parameters were derived to achieve robust hydrophobicity of a microstructured surface with hydrophobic and hydrophilic materials.

Effects of SO2 and NO on growth of Chlorella sp. KR-1.

Effects of the toxic compounds in flue gas, SOx and NOx, on growth of Chlorella sp. KR-1 have been determined. Although growth of KR-1 was suppressed by the toxic compounds, KR-1 exhibited excellent tolerances to SOx compared to other algal strains. When Chlorella KR-1 was cultured with the model gas containing 60 ppm SO2, the linear growth rate was 1.24 g/l day which is about 25% lower than that of the control culture aerated with the gas mixture containing no toxic compounds, SO2 and NO. KR-1 could grow even with the model gas containing 100 ppm SO2 and the linear growth rate of KR-1 in the culture was 0.78 g/l day. The period for lag phase was increased with increasing of SO2 concentration that also resulted in the decrease of the linear growth rate and the maximum cell concentration. Direct CO2 fixation by Chlorella KR-1 has been successfully done using actual flue gases from a liquified natural gas (LNG)- or diesel-fueled boiler. These results indicated that Chlorella KR-1 may be applied for direct CO2 fixation from actual flue gas.