- 著者
- Akita Hidetaka Ito Rie Kamiya Hiroyuki Kogure Kentaro Harashima Hideyoshi
- 出版者
- John Wiley & Sons
- 雑誌
- The Journal of Gene Medicine (ISSN:1099498X)
- 巻号頁・発行日
- vol.9, no.3, pp.197-207, 2007-03
- 被引用文献数
- 36
Background: Heterogeneity of transgene expression, the presence or absence (below the limit of detection) of transgene expression on a cell-by-cell basis, is a severe disadvantage in the use of cationic lipid-mediated gene vectors for gene therapy and experiments in molecular biology. Understandings of intracellular trafficking and the function (transgene expression) of vectors related to cellular physiology are essential in terms of clarifying the mechanism underlying the heterogeneity. Methods: To distinguish the contribution of nuclear transfer efficiency and subsequent intranuclear transcription efficiency to the overall heterogeneity in transgene expression, a novel imaging system was established for the dual visualization of the nuclear transfer of pDNA and marker gene expression (lacZ) in single cells. Results: The expression of LacZ occurred in only approximately 30% of HeLa cells of the nuclear pDNA-positive cells, indicating that intranuclear transcription efficiency contributed to the heterogeneity. Dual imaging against synchronized cells further revealed that the efficiency of nuclear delivery was comparable irrespective of cell cycle status, which is contrary to the generally accepted hypothesis that nuclear import of pDNA is enhanced during cell division when the nuclear membrane structure is perturbed. The most significant finding in the present study is that nuclear transcription efficiency in terms of the ratio of LacZ-positive cells to nuclear pDNA-positive cells drastically increased in the late S and G2/M phase. Conclusions: This is the first demonstration to show that cell cycle dependent intranuclear transcription appears to be responsible for the overall heterogeneity of transgene expression. Copyright © 2007 John Wiley & Sons, Ltd.
- 著者
- MAEBUCHI Motohiro SAMOTO Masahiko KOHNO Mitsutaka ITO Rie KOIKEDA Takashi HIROTSUKA Motohiko NAKABOU Yukihiro
- 出版者
- 公益社団法人 日本食品科学工学会
- 雑誌
- Food science and technology research (ISSN:13446606)
- 巻号頁・発行日
- vol.13, no.1, pp.45-53, 2007-02-01
- 参考文献数
- 34
- 被引用文献数
- 5 52
In this study, we compared the intestinal absorption from different sources with the same amino acid composition (protein, peptide, and amino acid mixture) by measuring changes in serum concentrations of various amino acids. Peptides used in this study were prepared by enzymatic digestion of 11S globulin (a major component of soy protein). Twelve normal male adults were asked to drink beverages containing 12.5g of the test sample, representing 5% suspension of each of the following three different preparations : 11S globulin, 11S peptide, and an amino acid mixture of equivalent composition. Venous blood samples were collected from 0 to 180min to measure free amino acid concentrations. The early slope of serum concentrations and values of area under the curves for each amino acid calculated from changes in serum amino acid concentrations were significantly greater for 11S peptide than for either 11S globulin or the amino acid mixture. This trend was especially remarkable for branched-chain amino acids and aromatic amino acids. Our results suggest that the ingestion of hydrolyzed soy protein results in faster and more efficient absorption than the consumption of protein or amino acid mixtures in healthy adult men.