Combination of cytomegalovirus enhancer with human cellular promoters for gene-induced chondrogenesis of human bone marrow mesenchymal stem cells

High-expression plasmid vectors for human mesenchymal stem cells (MSCs) were constructed by combination of cytomegalovirus immediate-early enhancer with cellular promoters. MSCs transfected with the vector showed higher transgene production of a cytokine, which increased the differentiation level to chondrocytes.     

Assessment of the antioxidant capacity of a fermented grain food product,Antioxidant Biofactor (AOB), by using pyranine and pyrogallol red as a combined probe

The role of antioxidants contained in foods, beverages, and supplements against oxidative stress has received much attention. The capacity of antioxidants has been assessed by various methods. In this study, the antioxidant capacity of a complex mixture of fermented grains has been assessed by using two probes, pyranine and pyrogallol red (PGR). A supplement commercialised as Antioxidant Biofactor, AOB, was used as a substrate. The extracts from AOB obtained with water and dimethylsulphoxide (DMSO) inhibited the free radical-induced consumption of pyranine and PGR in a concentration-dependent manner. They also suppressed the free radical-induced oxidation of human plasma. It was estimated that 1 g of AOB contained, as Trolox equivalent, roughly 0.13 and 0.24 mmol (2.5 and 4.7 wt%) antioxidants, which could be extracted by water and DMSO, respectively. This study shows that the combination of the above two probes is useful for assessing the total content and activity of antioxidants contained in complex mixtures.     

Toxicity and tetramine contents of salivary glands from carnivorous gastropods

Salivary glands from 29 species of marine carnivorous gastropods in nine families were examined for lethal activity against mice and tetramine content. Mouse lethality was assayed by intravenous injection of buffer extracts into mice, and was detected in 14 species. Heat-stability tests confirmed that toxins in four species were thermolabile, while those in eight species were thermostable. Based on the tetramine contents determined by the colorimetric method using methanolic extracts, the thermostable toxins in seven species (Neptunea eulimatalamellosa, N. vinosa, N. arthritica, N. bulbacea, N. intersculpta f. pribiloffensis, N. intersculpta f. frater pilsbry and Hemifusus tuba) were considered to be tetramine contained at high levels (more than 900 micrograms/g salivary gland), but that in one species (Buccinum opisthoplectum) appeared to be a low-molecular-weight compound differing from tetramine. It is interesting that one (Hemifusus tuba) of the seven species containing high amounts of tetramine belongs to the family Melongenidae, although the other six Neptunea species are members of the family Buccinidae, as expected from previous studies.     

Determination of sucralose in foods by HPLC using pre-column derivatization

The development of a sensitive pre-column derivatization high-performance liquid chromatography (HPLC) method for determination of sucralose is reported. Sucralose is converted into a strongly ultraviolet (UV)-absorbing derivative, possessing strong absorption at 260 nm, by treatment with p-nitrobenzoyl chloride (PNBCl). Homogenized samples were dialyzed and washed with a Bond Elut ENV cartridge, then the eluate was evaporated to dryness and the residue was derivatized. Subsequently, the sucralose derivative was purified with hexane-ethyl actate (9:1) in a silica cartridge, and then the sucralose derivative was eluted with acetone. HPLC was performed on a phenyl column, using acetonitrile-water (73:27) as a mobile phase with UV detection (260 nm). The calibration curve was linear in the range of 1 microgram/mL to 50 micrograms/mL of sucralose. The recoveries of sucralose from eight kinds of foods spiked at the levels of 0.20 and 0.05 g/kg of sucralose were more than 76.2% with SD values in the range from 0.90% to 4.31%. The quantitative limit of the developed method was 0.005 g/kg for sucralose in samples.     

Reliable indirect competitive ELISA used for a survey of ochratoxin A in green coffee from the North of Paraná State, Brazil.

The performance of an indirect competitive enzyme-linked immunosorbent assay (ic-ELISA) based on a monoclonal antibody (mAb) for ochratoxin A (OTA) detection was evaluated in a comparative study with high-performance liquid chromatography (HPLC) analysis using 68 freshly harvested coffee samples from the North of Paraná State, Brazil. The anti-OTA mAb showed high specificity and low cross-reactivity with OTA analogues (OTB and OTalpha), but cross-reacted with OTC. This ic-ELISA showed a detection limit of 3.75 ngg-1 sample, when compared to 0.80 ngg-1 by HPLC, with an ic-ELISA/HPLC correlation coefficient of 0.90. As regards OTA analysis of these coffee samples, natural contamination was detected in 10 samples (14.7%) by both methods, where the ic-ELISA values (range 3.9-7.3 ngg-1) were 1.1 to 1.6-fold higher than HPLC data (2.7-4.7 ngg-1). Five samples (7.4%) were OTA positive (range 0.84-1.30 ngg-1) only by HPLC assay, probably due to the higher detection limit reached by ic-ELISA. OTA was undetectable in 53 samples (77.9%) by both methods, while all positive samples (range 0.84-7.30 ngg-1) showed OTA levels lower than 8 ngg-1 (maximum limit recommended by the European Union). The matrix interference of green coffee was minimized by dilution of sample extracts before carrying out the ELISA assay. This mAb-based ic-ELISA can be effectively applied for OTA screening in coffee, because it is simple, sensitive and sample preparation is easy.     

Characteristics of neutralization of acids by newly isolated fungal cells

Soil microorganisms play an important role in maintaining soil pH at levels suitable for other soil organisms. To clarify the biological neutralization mechanism in soil, we isolated soil microorganisms showing a high ability to neutralize acids and studied their characteristics. From our taxonomic study, three isolated strains were identified as filamentous fungi, namely Mucor sp., Aspergillus fumigatus, and Aureobasidium pullulans. These strains could secrete basic materials, such as ammonia, for neutralization, grow in the medium at pH 4.0 and increase the pH of the medium to approximately 8.0. These microbial cells could neutralize not only nitric acid but also sulfuric and hydrochloric acids. The strains could also grow by utilizing nitric acid as a sole nitrogen source. In the soil containing these organisms, the pH was maintained in the neutral range by the buffering action of basic materials that they secrete. These results suggest that these fungal cells are useful for protecting the soil from acidification by acid rain.     

Memory encoding by theta phase precession in the hippocampal network

Recent experimental evidence on spike-timing-dependent plasticity and on phase precession (i.e., the theta rhythm dependent firing of rat hippocampal cells) associates the contribution of phase precession to episodic memory. This article aims at clarifying the role of phase precession in memory storage. Computer simulations show that the memory storage in the behavioral timescale varies in timescale of the temporal sequence from half a second to several seconds. In contrast, the memory storage caused by traditional rate coding is restricted to the temporal sequence within 40 ms. During phase precession, memory storage of a single trial experience is possible, even in the presence of noise. It is therefore concluded that encoding by phase precession is appropriate for memory storage of the temporal sequence in the behavioral timescale.     

Effects of rice cleaning and cooking process on the residues of flutolanil,fenobucarb, silafluofen and buprofezin in rice

We studied the effect of cleaning and cooking on the residues of flutolanil, fenobucarb, silafluofen and buprofezin in rice. The rice had been sprayed in a paddy field in Wakayama city, with 3 kinds of pesticide application protocols: spraying once at the usual concentration of pesticides, repeated spraying (3 times) with the usual concentration of pesticides and spraying once with 3 times the usual concentration of pesticides. The residue levels of pesticide decreased during the rice cleaning process. Silafluofen, which has a higher log Pow value, remained in the hull of the rice. Fenobucarb, which has a lower log Pow value, penetrated inside the rice. The residue concentration of pesticide in polished rice was higher than that in pre-washed rice processed ready for cooking. During the cooking procedure, the reduction of pesticides in polished rice was higher than that in brown rice.     

An 8.1-ns Column-Access 1.6-Gb/s/pin DDR3 SDRAM With an 8:4 Multiplexed Data-Transfer Scheme

Three circuit techniques for an 8.1-ns column-access 1.6-Gb/s/pin 512-Mb DDR3 SDRAM using 90-nm dual-gate CMOS technology were developed. First, an 8:4 multiplexed data-transfer scheme, which operates in a quasi-4-bit prefetch mode, achieves a 3.17-ns reduction in column-access time, i.e., from 11.3 to 8.13 ns. Second, a dual-clock latency counter reduces standby power by 22% and cycle time from 1.7 to 1.2 ns. Third, a multiple-ODT-merged output buffer enables selection of five effective-resistance values Rtt (20, 30, 40, 60, and 120 Omega) without increasing I/O capacitance. Based on these techniques, 1.6-Gb/s/pin operation with a 1.36-V power supply and a column latency of 7 was accomplished