The Ocular Protective Effects of Nano/Submicron Particles Prepared from Lycium barbarum Fruits Against Oxidative Stress in an Animal Model.

Purpose: To investigate the antioxidative properties of Lycium barbarum (LB) fruits in the eyes and to study whether LB fruits prepared with new nanotechnology have stronger antioxidative effects. Methods: Fourteen days post-supplementation with milled or blended LB fruits, intravitreal paraquat (PQ) was injected into Wistar rats to create oxidative stress. After an additional 14-day supplementation with LB fruits, the rats were sacrificed. An electroretinogram (ERG) was performed to evaluate retinal function before and after the PQ injection. Expression levels of antioxidative responders' mRNA in retina were detected by reverse transcription-polymerase chain reaction. Superoxide dismutase (SOD) and glutathione reductase activity in the aqueous humor (AqH) were analyzed by ELISA. Immunohistochemistry was conducted to evaluate the morphological changes of retina and the levels of oxidative biomarkers. The levels of cell apoptosis were assessed by the terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) assay. The reactive oxygen species (ROS) levels in AqH were measured by chemiluminescence methods. Results: The murine eyes supplemented with LB fruits exhibited several changes compared with the control group. The ERGs revealed significant improvement in retinal function. The mRNA expression levels of oxidative responders were downregulated in the retinas. The ROS was significantly reduced in the retinas, but the SOD meaningfully increased in the AqH. Immunohistochemistry staining and TUNEL assays showed decreased incidences of oxidative biomarkers and apoptosis in the retinas. Milled LB fruits exhibited better antioxidative effects than blended fruits. Conclusions: Milled LB fruits demonstrated superior protection against oxidative threats than blended fruits. Thus, these fruits could be an inexpensive supplement for many oxidative stress-related ocular diseases.

Enhancing cellulose functionalities by size reduction using media-mill.

This study explored the feasibility of enhancing cellulose functionalities by using media milling to reduce the size of cellulose particles, and assayed various physicochemical and physiological properties of the resulting cellulose. Cellulose has been recognized as dietary fiber by USFDA due to its health benefits. However, its properties like low degradability, stiff texture, and insolubility in water limits its applicability in foods. Milling reduced the volume mean size of cellulose from 25.7 µm to 0.9 µm, which in turn increased the specific surface area (36.78-fold), and swelling capacity (9-fold). Conversely, a reduction in the bulk density (1.41 to 1.32 g/mL) and intrinsic viscosity (165.64 to 77.28 mL/g) were found. The milled cellulose also had significantly enhanced capacity for holding water and binding bile acids and sugars. Moreover, the size reduction also resulted in increased fermentability of cellulose into short chain fatty acids using three human fecal microflora samples. The increase in production of acetate (2880.60%), propionate (2738.52%), and butyrate (2865.89%) after fermentation of cellulose for 24 h were significantly enhanced by size reduction. With these improved characteristics, the milled cellulose might have beneficial physiological effects including laxation as well as reduced blood cholesterol and glucose attenuation.

Preparation of nano/submicrometer Ganoderma tsugae and its mutagenic potencies and cytotoxicity.

This study explored the feasibility of preparing nano/submicrometer particles from Ganoderma tsugae to enhance the contents of bioactive compounds and to assess its mutagenic potencies and cytotoxicity. Hot-water extract, a common product, was employed as a reference. After 3 h of media milling, almost all of the particles were smaller than 1 µm with a number-mean diameter of 0.11 µm. There were about 62% particles smaller than 0.1 µm in terms of number of particles. Scanning electron microscopy (SEM) confirmed the presence of particles at nano/submicrometer scale. The content of 1→3-ß-D-glucan in nano/submicrometer G. tsugae was 3.5 times of that in hot-water extract. Both nano/submicrometer and hot-water extract G. tsugae exhibited no mutagenic potential to Salmonella Typhimurium tester strains. Cell toxicity test also confirmed the safety of both nano/submicrometer and hot-water extract G. tsugae. The effect of media milling on the structural change of hyphae was also discussed.

Preparation and physicochemical properties of whole-bean soymilk.

Whole-bean soymilk has been prepared by using media-milling. Some characteristics of media-milled soymilk have been determined and compared with filtered soymilk (similar to commercial ones) and whole-bean soymilk prepared by blending. There existed particles in the nano/submicrometer scale in both media-milled and filtered soymilk. The particles in blended soymilk were greater than 1 µm. Media-milled soymilk was the most stable among three samples, even after autoclaving. Solid recovery (98.44 ± 0.16%), viscosity (160.59 ± 4.26 cps), dietary fiber (22.68 ± 0.97% on dry basis), total polyphenol recovery (95.15 ± 7.09%), and isoflavone content (4.42 ± 0.03 mg/g dry solid) of media-milled samples were greater than those of filtered ones. Aglycones, the most bioactive form of isoflavone, in autoclaved media-milled soymilk were more than 2-fold those in autoclaved filtered soymilk. With almost no okara generated, the media-milled soymilk retained fiber in soybeans which would be beneficial to human health.

Preparation of nano/submicrometer yam and its benefits on collagen secretion from skin fibroblast cells.

Nano/submicrometer-scaled yam particles have been prepared by using media-milling. The particle size of media-milled yam was confirmed by the laser light scattering method and scanning electron microscopy. Influences of media-milled yam on skin fibroblast cells (WS1) were evaluated. The size reduction did not significantly alter the proximate composition, and the presence of nanoparticles was not toxic to WS1 cells. The contents of bioactive compounds (diosgenin, stigmasterol, and ß-sitosterol) were significantly increased by media-milling, which enhanced the secretion of hTGF-ß and inhibited the formation of MMP-1. Thus, the collagen secretion from WS1 was significantly increased by size reduction. Diosgenin was employed as a positive control. Nevertheless, media-milled yam exhibited greater effects on WS1 cells than diosgenin. It appeared that both diosgenin and size reduction were helpful for enhancing the secretion of collagen by WS1 cells. In addition, the irritancy of yam was eliminated by media-milling.

Acrylamide content distribution and possible alternative ingredients for snack foods.

Acrylamide (AA) contents in 294 snack foods including cereal-based, root- and tuber-based, and seafood-based foods, nuts, dried beans, and dried fruits purchased in Taiwan were determined by gas chromatography-mass spectrometry in this study. The highest levels of average AA content were found in root- and tuber-based snack foods (435 µg/kg), followed by cereal-based snack foods (299 µg/kg). Rice flour-based, seafood-based, and dried fruit snack foods had the lowest average AA content (<50 µg/kg). This is the first large surveillance of AA content in snack foods in Taiwan. The results could provide important data regarding intake information from the snack foods. In addition, the results showed a great diversity of AA content in snack foods prepared from different ingredients. Rice- and seafood-based products had much lower AA than those made from other ingredients. This information could constitute a good reference for consumers to select products for healthy snacking.

Effect of Goji (Lycium barbarum) on expression of genes related to cell survival.

This study investigated the interrelationship between Lycium barbarum (goji) and gene expression in mouse spleen. Oligomicroarray technology was employed to explore the comprehensive response of gene expression and to screen candidate marker genes in the spleens of mice fed a goji suspension. Goji was micronized by media milling and then used to evaluate the effect of size reduction. The average diameter of nano/submicrometer goji was about 100 nm, which exhibited no cytotoxicity to cell lines IEC-6 (rat normal small intestinal cell line) and Caco-2 (human colon adenocarcinoma cell line). It was found that three genes, TNF, Nfkb1, and Bcl-2, were up-regulated and two genes, APAF-1 and caspase-3, were down-regulated by goji. This phenomenon could be helpful for cytoprotection when cells undergo stress or damage that induces the apoptotic pathway. Size reduction into nano/submicrometer scale enhanced bioactivity.

Physico-chemical characteristics of media-milled corn starch.

Nano/submicrometer scale corn starch particles have been prepared by using media milling. Molecular weight and pasting properties of milled starch were determined by HPSEC-MALLS-RI and an oscillatory rheometer, respectively. After 30-min of milling, the number-average diameter of the starch particle was reduced from 9.61 to 0.26 µm. Particle size was further reduced by extended milling. Microscopic observation confirmed the presence of particles smaller than 100 nm. As starch granules were being disintegrated, the mechanical energy imparted by media milling resulted in 43.7% of starch damage and 55% degree of gelatinization during the initial 30 min. In the meanwhile, the weight average molecular weight was reduced from 9.98 to 7.63 × 10(6) g/mol. The pasting properties were significantly altered by milling, which resulted in lower onset and peak temperature, and reduction in breakdown and set back. The importance of molecular weight on the physicochemical properties of milled starch was discussed.

Differential tissue distribution of sesaminol triglucoside and its metabolites in rats fed with lignan glycosides from sesame meal with or without nano/submicrosizing.

Lignan glycosides are important functional compounds in sesame meal. In the present study, we investigated whether the tissue distribution of nano/submicrosized lignan glycosides from sesame meal (N-LGSM) differs from lignan glycosides from sesame meal (LGSM). LGSM was nano/submicrosized with 0.3 mm zirconia beads as the milling media. The average particle size of the 4% LGSM aqueous suspension reduced rapidly from approximately 2 microm to 200 nm after media milling at an agitation speed of 3600 rpm for 30 min. We examined the tissue distribution of sesaminol triglucoside (ST), the main component in LGSM, in Sprague-Dawley (SD) rats. The concentrations of ST were determined in various tissues and plasma within a 24 h period after oral administration of N-LGSM and LGSM (800 mg/kg of body weight). The results showed that higher concentrations of ST and its metabolites (sesaminol, sesaminol sulfate, and sesaminol glucuronide) were found in N-LGSM compared to those in LGSM in most tissues, especially liver and small intestine. Sesaminol glucuronide was the main metabolite in rats. After 3 h of oral administration, around 70% higher concentration of sesaminol glucuronide was found in N-LGSM compared to that in LGSM. This study clearly showed that LGSM is more bioavailable after nano/submicrosizing.