Detection of genetically modified organisms in foreign-made processed foods containing corn and potato

Investigations of the validity of labeling regarding genetically modified (GM) products were conducted using polymerase chain reaction (PCR) methods for foreign-made processed foods made from corn and potato purchased in the Tokyo area and in the USA. Several kinds of GM crops were detected in 12 of 32 samples of processed corn samples. More than two GM events for which safety reviews have been completed in Japan were simultaneously detected in 10 samples. GM events MON810 and Bt11 were most frequently detected in the samples by qualitative PCR methods. MON810 was detected in 11 of the 12 samples, and Bt11 was detected in 6 of the 12 samples. In addition, Roundup Ready soy was detected in one of the 12 samples. On the other hand, CBH351, for which the safety assessment was withdrawn in Japan, was not detected in any of the 12 samples. A trial quantitative analysis was performed on six of the GM maize qualitatively positive samples. The estimated amounts of GM maize in these samples ranged from 0.2 to 2.8%, except for one sample, which contained 24.1%. For this sample, the total amount found by event-specific quantitative analysis was 23.8%. Additionally, Roundup Ready soy was detected in one sample of 21 potato-processed foods, although GM potatoes were not detected in any sample.     

Properties of cold-rolled high-strength steel sheets

For high-strength steel sheets, a new concept has become necessary,viz, the relation between strength and formability. When the relation between tensile strength and elongation is appraised for ranking in terms of the strengthening mechanism, it is found that the substitutional solid-solution hardening type is excellent and the precipitation hardening type is inferior. In batch annealing, the phosphorus-added aluminum-killed steel sheets are representative of the former type having excellent formability as indicated by a highr values despite their tensile strength of 450 N/mm². The titanium-added aluminum-killed steel sheets, which are representative of the latter type, have a tensile strength of 600 N/mm² and a relatively highr value. Continuous annealing of the highstrength steel sheets of the same chemical composition causes higher yield stresses and combinations of yield stress and elongation. Alternatively with continuous annealing the same level of strength can be achieved with smaller additions of alloying elements than with batch annealing. An additional advantage of continuous annealing is the uniformity of properties along the length of the coil. The rapid cooling possible after continuous annealing allows production of high strength steel sheets having excellent mechanical properties that are unobtainable in the batched annealed steels. For example, steel sheets of 0.4 pet Si and 1.4 pet Mn after continuous annealing, haven values and Erichsen values as higher than conventional low-carbon rimmed or capped steel sheets even while they have a tensile strengths of 550 N/mm².     

Sweeteners interacting with the transmembrane domain of the human sweet-taste receptor induce sweet-taste synergisms in binary mixtures

Sweet enhancing effect of neohesperidin dihydrochalcone (NHDC) or cyclamate has been reported to be synergistic in human sensory tests. However, little is known about whether these synergisms are caused by the mechanism mediated by the human sweet-taste receptor. Here, we examined the sweetness intensity of sweet tastant mixtures by measuring the responses of cultured cells stably expressing the human sweet-taste receptor. The results showed that the cell response to sucrose was synergistically potentiated by the addition of NHDC or cyclamate. Moreover, a point mutation in the transmembrane domain of hT1R3 almost completely eliminated the enhancing effects of NHDC and cyclamate. These results suggest that ligand–receptor interactions in the transmembrane domain of hT1R3 are necessary for NHDC and cyclamate to elicit the synergistic potentiation of the receptor activation. Our results may provide the foundation of a molecular basis for receptor-based synergisms of sweet tastes in mixtures of diverse sweet substances.     

Comparison of isolation methods using commercially available kits for obtaining extracellular vesicles from cow milk

Extracellular vesicles (EV) are important for delivering biologically active substances to facilitate cell-to-cell communication. Milk-derived EV are widely known because of their potential for immune enhancement. However, procedures for isolating milk-derived EV have not been fully established. To obtain pure milk-derived EV and accurately reveal their function, such procedures must be established. The aim of the present study was to compare methods using commercially available kits for isolating milk-derived EV. Initially, we investigated procedures to remove casein, which is the major obstacle in determining milk-derived EV purity. We separated whey using centrifugation only, acetic acid precipitation, and EDTA precipitation. Then, we isolated milk-derived EV by ultracentrifugation, membrane affinity column, size exclusion chromatography (SEC), polymer-based isolation, or phosphatidylserine-affinity isolation. Using EV count per milligram of protein, which is a good indicator of purity, we determined that acetic acid precipitation was the best method for removing casein. Using nanoparticle tracking analysis, protein quantity analysis, and RNA quantity analysis, we comprehensively compared each isolation method for its purity and yield. We found that SEC-based qEV column (Izon Science) could collect purer milk-derived EV at higher quantities. Thus, a combination of acetic acid precipitation and qEV can effectively isolate high amounts of pure extracellular vesicles from bovine milk.     

Levels of Vibrio parahaemolyticus and thermostable direct hemolysin gene-positive organisms in retail seafood determined by the most probable number-polymerase chain reaction (MPN-PCR) method

The incidence and levels of Vibrio parahaemolyticus and thermostable direct hemolysin gene (tdh)-positive organisms in retail seafood were determined. The most probable number-polymerase chain reaction (MPN-PCR) method using a PCR procedure targeting the species-specific thermolabile hemolysin gene (tlh) and tdh was used to determine the levels of V. parahaemolyticus and tdh-positive organisms, respectively. In seafood for raw consumption, V. parahaemolyticus was found in four (13.3%) of 30 fish samples, 11 (55.0%) of 20 crustacean samples, and 29 (96.7%) of 30 mollusc samples. Levels of V. parahaemolyticus were below 10(4) MPN/100 g in all fish and crustacean samples tested. However, they were above 10(4) MPN/100 g in 11 (36.7%) of the 30 mollusc samples. In all seafood for raw consumption, the level of tdh-positive organisms was below the limit of detection (< 30 MPN/100 g). In seafood for cooking, V. parahaemolyticus was found in 15 (75.0%) of 20 fish samples, nine (45.0%) of 20 crustacean sample, and 20 (100%) of 20 mollusc samples. Levels of V. parahaemolyticus were above 10(4) MPN/100 g in only three (15.0%) and one (5.0%) of the 20 fish and 20 crustacean samples, respectively. However, they were above 10(4) MPN/100 g in 18 (90.0%) of the 20 mollusc samples. In seven (35.0%) of the 20 mollusc samples, tdh-positive organisms were found and their levels ranged from 3.6x10 to 1.1 x 103 MPN/100 g. From four of seven tdhpositive samples, tdh-positive V. parahaemolyticus was isolated.