Soybean ( Glycine max [L.] Merr.) seed is about 40% protein, most of which is accounted for by the major storage proteins β-conglycinin (7S globulin) and glycinin (11S globulin). β-conglycinin, which consists of three subunits, α, α', and β, is the main allergen in soybean. Accordingly, elimination of β-conglycinin from seed is one of the goals of soybean breeding programs. Soybean accessions PI 200485 and CS 1150 lack the α'- and α-subunits of β-conglycinin, respectively. In this study, we developed a 3-betacon marker that could simultaneously identify genotypes for the three subunit genes of β-conglycinin. Interestingly, the three subunit genes were amplified with three distinct bands representing varied amplicon sizes. Mutant alleles could be identified in progeny of a three-way cross, and the corresponding α- and α'-subunit proteins were clearly absent in each mutant accession. 3-betacon marker will be highly useful for soybean breeding programs to develop soybean cultivars with decreased β-conglycinin contents. The online version of this article (doi: 10.5073/JABFQ.2015.088.041) contains supplementary files .
Linear viscoelastic analysis is performed to predict residual stresses occurred in composites during cure. Viscoelastic constitutive equations are defined as functions of temperature and degree of cure. An algorithm of curing process is developed to reduce curing time using constraints imposed on the temperature, the degree of cure and the residual stresses. An optimal cure cycle determined by this curing process algorithm is compared with MRC (manufacturer's recommended we cycle). Since the viscoelastic residual stresses are mostly relaxed at curing stage and hardly relaxed at cool-down stage, there is no big difference between final residual stresses calculated by the optimal cure cycle and the MRC. But the proposed cure cycle in this paper shows a possibility to reduce the curing time considerably as compared to the MRC.
Stevioside, a natural sweeteners presently used in various kinds of food and food products in Korea, was evaluated for its toxicity potential in the 14 day feeding study using B6C3F1 mice. Stevioside was added to the diet at different concentrations of 0.31, 0.62, 1.25, 2.5 and 5%, and was administered for 14 consecutive days. An increase of liver organ weight in male mice was observed. No diet-related differences were noted in clinical signs, food consumption, and gross and histopatholgical evaluation. Based on these results, we concluded that the concentration of 5% in the diet was a suitable maximum tolerable dose of stevioside for a 90 day study in mice.
Over the past several years, viral and non-viral gene delivery systems have been intensively developed to establish an ideal delivery vector for cancer gene therapy. Among the large variety of virus vectors currently being developed, the vector based on recombinant AAV (rAAV) is one of those that are closest to the ideal vector due to the following features; the lack of pathogenicity and toxicity, ability to infect dividing and non-dividing cells of various tissue origins, a very limited host immune response and long-term persistent expression. However, there is concern that two major obstacles of AAV serotype 2 (AAV2), the relative paucity of AAV receptors on certain cell types and the presence of pre-existing immunity to AAV2 in humans might limit its clinical applications in humans. To overcome these limitations, we first investigated in vitro transduction efficiencies for 7 different AAV serotypes (AAV1 – AAV6, AAV8) in 12 human tumor cell lines (MKN74, AGS, NCI-H460, HT1080, HepG2, SK-Hep1, HCT-116, 5637, UMUC3, HeLa, DU145 and MCF7). We employed "pseudotyped" AAV vector systems, in which rAAV2 vector genomes containing the CMV promoter/ enhancer, the green fluorescent protein (GFP) cDNA and SV40 poly (A) signal, flanked by AAV2 ITR sequences are cross-packaged into the capsid proteins of the other AAV serotypes (several plasmids are kindly provided by Dr. Katherine A. High, University of Pennsylvania Medical Center). As we reported previously, the efficiency of rAAV2 infection into most of the tested cancer cell lines was significantly high (>90%), whereas the transduction efficiencies for other 5 different rAAV serotypes were varied and greatly low. On the other hand the efficiency of rAAV5 transduction in various tumor cell lines were comparably high, mostly similar to that of rAAV2. Using the xenograft nude mice implanted with either NCI-H460 or HCT-116 cancer cells, we also studied in vivo transduction efficiency for several pseudotyped rAAV vectors, which were shown to have a host range different from rAAV2. Taken altogether our results suggest that the pseudotyped rAAV5 vector can be developed as one of the most efficient gene delivery systems for cancer gene therapy to overcome the anti-AAV2 immune response. (This study was supported by the grant from the Ministry of Science and Technology (M10534040005-05N3404-00511), Seoul, and the Chung-Buk Pioneering Bioindustry R&D Grant, Chung-Buk, Korea.)
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