Antioxidant behaviour of carotenoids highly accumulated in HepG2 cells

The antioxidant behaviour of major dietary carotenoids accumulated at high concentrations in human hepatoma HepG2 cells was evaluated, in comparison with α-tocopherol. The cells that accumulated carotenoids and α-tocopherol at levels higher than the values reported in the human liver were exposed to mild oxidative stress with tert-butylhydroperoxide. β-Carotene (>2.6 nmol/mg protein) and astaxanthin (>1.8 nmol/mg protein) significantly suppressed lipid peroxidation, while β-cryptoxanthin and lutein did not. α-Tocopherol remarkably suppressed lipid peroxidation with an IC₅₀ value of 0.16 nmol/mg protein. Neither α-tocopherol nor any of the carotenoids except for lycopene showed pro-oxidant action even at high cellular concentrations. The antioxidant behaviours of carotenoids in a cellular milieu were quite different from those previously found in liposomes and homogeneous solutions. Further studies are required to assess the implications of the antioxidant behaviours found in the cultured cells on human health.     

Ultrastructural analysis of buckwheat starch components using atomic force microscopy

Morphological and structural features of buckwheat starch granules and nanocrystals were examined using atomic force microscopy and dynamic light scattering. Partially digested starch granules revealed a clear pattern of growth rings with the central core revealing lamellar structure. Atomic force microscopy and dynamic light scattering experiments revealed that the buckwheat starch granules were polygonal in shape and were in the range of 2 to 19 μm in diameter. The optimized acid hydrolysis process produced nanocrystals with the shape of spherical structure with lengths ranging from 120 to 200 nm, and the diameter from 4 to 30 nm from aqueous suspensions of buckwheat starch solution. The sorption isotherms on buckwheat starch nanocrystal/glycerol composite exhibited a 3-stage transition of moisture in the blending. The biocompatible nature of buckwheat starch nanocrystals and their structural properties make them a promising green nanocomposite material. PRACTICAL APPLICATION: Buckwheat starches had never been studied on a nanoscale, but we have achieved new understanding of starch granule morphology and concentric growth rings using nanoscale imaging. Since buckwheat is an underutilized crop, we foresee the potential application of buckwheat starch, starch-based nanocrystals, and nanoparticles, to expand markets and encourage producers to expand their buckwheat acreage. The atomic force image analysis suggests that buckwheat starch could be used as a new biopolymer material in food industries.     

Isolation of methyl syringate as a specific aflatoxin production inhibitor from the essential oil of Betula alba and aflatoxin production inhibitory activities of its related compounds

Methyl syringate was isolated from the essential oil of Betula alba as an aflatoxin production inhibitor. It inhibited aflatoxin production of Aspergillus parasiticus and Aspergillus flavus with IC₅₀ values of 0.9 and 0.8 mM, respectively, without significantly inhibiting fungal growth. Methyl syringate reduced mRNA levels of genes (aflR, pksA, and omtA) encoding proteins required for aflatoxin biosynthesis. Methyl gallate, methyl 3,4,5-trimethoxybenzoate, and methyl 3-O-methylgallate inhibited both aflatoxin production and fungal growth of A. parasiticus and A. flavus. However, their acids and syringic acid did not inhibit aflatoxin production and growth of A. parasiticus significantly, although gallic acid inhibited aflatoxin production of A. flavus with selectivity. The 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging activity of methyl syringate was much weaker than that of gallic acid. These results showed that methyl syringate has a unique inhibitory activity toward aflatoxin production with a different mode of action from that of gallic acid.     

Characterization and gene expression profiles of thermotolerant Saccharomyces cerevisiae isolates from Thai fruits

For industrial applications, fermentation of ethanol at high temperature offers advantages such as reduction in cooling costs, reduced risk of microbial contamination and higher efficiency of fermentation processes including saccharification and continuous ethanol stripping. Three thermotolerant Saccharomyces cerevisiae isolates (C3723, C3751 and C3867) from Thai fruits were capable of growing and producing 38 g/L ethanol up to 41°C. Based on genetic analyses, these isolates were prototrophic and homothallic, with dominant homothallic and thermotolerant phenotypes. After short-term (30 min) and long-term (12 h) exposure at 37°C, expression levels increased for the heat stress-response genes HSP26, SSA4, HSP82, and HSP104 encoding the heat shock proteins small HSP, HSP70, HSP90 and the HSP100 family, respectively. In isolates C3723 and C3867, expression was significantly higher than that in reference isolates W303 and TISTR5606 for TPS1 encoding trehalose-6-phosphate synthase, NTH1 encoding neutral trehalase and GSY1 encoding glycogen synthase. The results suggested that continuous high expression of heat stress-response genes was important for the long-term, heat stress tolerance of these thermotolerant isolates.     

Fates of foodborne pathogens in raw hams manufactured rapidly using a new patented method

To manufacture raw ham in an efficient manner, we recently developed a new system in which presliced pork loin was used, and the processing time was reduced to 5% of the conventional method. This study aimed to examine whether this raw ham could be as safe as ham produced by the conventional method. Pork loin spiked with enterohemorrhagic Escherichia coli serotype O157:H7, Listeria monocytogenes serotype 1/2c, Salmonella enterica serovar Enteritidis, and Staphylococcus aureus were processed using either the new or conventional method. The fate of the foodborne pathogens and behavior of hygiene indicator bacteria were examined. Whereas nitrite had disappeared during the conventional packaging process, the reduced processing time in the new system allowed for the ham to be vacuum packed with retention of the nitrite (6.9±1.2 ppm, P<0.01). This accounts for the prominent decrease in L. monocytogenes (2.3 log reduction in 35 days) and S. aureus (3.3 log reduction in 13 days) counts during storage. E. coli O157 and Salmonella Enteritidis were likely resistant to the nitrite in the ham. However, they were unable to multiply in the ham and decreased gradually as in the conventionally produced ham. The bacteriostatic nature of the raw ham was also indicated by the gradual decrease in coliforms (1.3 log reduction in 13 days) in nonspiked ham. In conclusion, the raw ham produced using presliced pork loin is practically as safe as conventionally produced raw ham. It is worth validating these results in a small-scale production setting.     

Synthesis and biological properties of sequence-specific DNA-alkylating pyrrole-imidazole polyamides

In recent years, many diseases including cancer and hereditary and viral diseases have been understood at the DNA sequence level. Direct control of the expression level of a specific gene would provide a promising approach for knowledge-based therapy. N-Methylpyrrole (Py) and N-methylimidazole (Im) polyamides are a new type of small compound that precisely bind to the minor groove of the DNA duplex in a sequence-specific fashion and recruit alkylating agents to the target sequence. We designed and synthesized a series of sequence-specific alkylating Py-Im polyamide conjugates that selectively alkylate predetermined DNA sequences. We have demonstrated that sequence-specific alkylating agents possess gene-silencing activities and a promising potency against human cancer cell lines as well as against xenografts of human cancer cell lines. In this Account, we focus on recent progress in alkylating Py-Im polyamides with regard to sequence specificity and biological activities and the future direction of rational molecular design of genetic switches in the postgenome era is described.     

Preparation of ultrathin self-standing polyelectrolyte multilayer membranes at physiological conditions using pH-responsive film segments as sacrificial layers

A new system to obtain ultrathin self-standing polyelectrolyte multilayer membranes at physiological conditions is introduced. On the surface of a substrate, a hybrid film structure composed of two compartments, (1) a pH-responsive film segment formed via hydrogen bonds and (2) a polyelectrolyte multilayer film on top of 1, was assembled. The pH-responsive polymer multilayer segments disintegrate at a neutral pH and release self-standing polyelectrolyte multilayer films. The obtained self-supporting polyelectrolyte multilayer membranes had thicknesses of 55 to several hundred nanometers and areas of a few square centimeters, approximately. The preparation method introduced here avoids harsh release conditions and thus broadens the choice of materials that can be incorporated into the self-standing film.     

Single-shot surface profiling by local model fitting

A new surface profiling algorithm called the local model fitting (LMF) method is proposed. LMF is a single-shot method that employs only a single image, so it is fast and robust against vibration. LMF does not require a conventional assumption of smoothness of the target surface in a band-limit sense, but we instead assume that the target surface is locally constant. This enables us to recover sharp edges on the surface. LMF employs only local image data, so objects covered with heterogeneous materials can also be measured. The LMF algorithm is simple to implement and is efficient in computation. Experimental results showed that the proposed LMF method works very well.     

Advanced SOI p-MOSFETs with strained-Si channel onSiGe-on-insulator substrate fabricated by SIMOX technology

We have newly developed an advanced SOI p-MOSFET with strained-Si channel on insulator (strained-SOI) structure fabricated by SIMOX (separation-by-implanted-oxygen) technology. The characteristics of this strained-SOI substrate and electrical properties of strained-SOI MOSFETs have been experimentally studied. Using strained-Si/relaxed-SiGe epitaxy technology and usual SIMOX process, we have successfully formed the layered structure of fully-strained-Si (20 nm)/fully-relaxed-SiGe film (290 nm) on uniform buried oxide layer (85 nm) inside SiGe layer. Good drain current characteristics have been obtained in strained-SOI MOSFETs. It is found that the hole mobility is enhanced in strained-SOI p-MOSFETs, compared to the universal hole mobility in an inversion layer and the mobility of control SOI p-MOSFETs. The enhancement of the drive current has been kept constant down to 0.3 μm of the effective channel length