Direct immunological identification of full-length cDNA clones for plant protein without gene fusion to E. coli protein

AbstractBy immunological screening of a cDNA library constructed from potato tuber poly(A)+ RNA and Escherichia coli expression vector pUC8 by the vector-primer and linker procedure of Okayama and Berg [(1982) Mol. Cell Biol. 2, 161-170], nearly full-length cDNA clones for patatin, a major protein of potato tuber, were identified. The cDNA carrying part of the 5'-noncoding region of the patatin mRNA, in addition to entire coding and 3'-noncoding regions, expressed prepatatin in E. coli cells by translational initiation inside cDNA. These results suggest that nearly full-length cDNA clones with entire coding region can be identified directly by immunological screening without gene fusion to E. coli proteins at least for some plant mRNAs

Cell-free synthesis of alkaline lipase, a glyoxysomal membrane protein, from castor bean endosperm

AbstractPolyadenylated RNA from castor bean endosperm was translated in a wheat germ cell-free protein synthesis system, and alkaline lipase, an integral glyoxysomal membrane protein, was immunoprecipitated. The apparent molecular mass of the lipase synthesized in vitro was slightly higher than that of the mature enzyme (62 kDa). When mRNA derived from free and membrane-bound polysomes in the endosperms was translated in vitro, the lipase was predominantly recovered from the products of the free polysomal mRNA, suggesting that the membrane protein is post-translationally inserted into the membrane. The amino-terminal 8 amino acid residues of the mature lipase were sequenced

A circulating subset of iNKT cells mediates antitumor and antiviral immunity

新規の循環型iNKT細胞を発見 --抗腫瘍・抗ウイルス感染効果の高い免疫細胞療法の開発への貢献に期待--. 京都大学プレスリリース. 2022-10-24.Invariant natural killer T (iNKT) cells are a group of innate-like T lymphocytes that recognize lipid antigens. They are supposed to be tissue resident and important for systemic and local immune regulation. To investigate the heterogeneity of iNKT cells, we recharacterized iNKT cells in the thymus and peripheral tissues. iNKT cells in the thymus were divided into three subpopulations by the expression of the natural killer cell receptor CD244 and the chemokine receptor CXCR6 and designated as C0 (CD244⁻CXCR6⁻), C1 (CD244⁻CXCR6⁺), or C2 (CD244⁺CXCR6⁺) iNKT cells. The development and maturation of C2 iNKT cells from C0 iNKT cells strictly depended on IL-15 produced by thymic epithelial cells. C2 iNKT cells expressed high levels of IFN-γ and granzymes and exhibited more NK cell–like features, whereas C1 iNKT cells showed more T cell–like characteristics. C2 iNKT cells were influenced by the microbiome and aging and suppressed the expression of the autoimmune regulator AIRE in the thymus. In peripheral tissues, C2 iNKT cells were circulating that were distinct from conventional tissue-resident C1 iNKT cells. Functionally, C2 iNKT cells protected mice from the tumor metastasis of melanoma cells by enhancing antitumor immunity and promoted antiviral immune responses against influenza virus infection. Furthermore, we identified human CD244⁺CXCR6⁺ iNKT cells with high cytotoxic properties as a counterpart of mouse C2 iNKT cells. Thus, this study reveals a circulating subset of iNKT cells with NK cell–like properties distinct from conventional tissue-resident iNKT cells

People and communities in modern cities

This study is divided into five chapters plus an. introduction which explains the main. objective of this thesis. The first chapter is an urban-social development of the origin of the city until modern times. This development describes the crises, chaos and recovery of cities. It also explains the process of growths and changes throughout history. Every new technological improvement brought urban and social changes in the city which were expressed with spatial features. The second chapter is related to the ecological pattern of the city and its influence over the total structure of the city, community, and the people. This influence is remarkable in all periods in the history of urban and rural development, without exception. The third chapter is a study of the American cities, that evolves from the early colonial times until today. It explains its growth and development and how the United States became, economically, the most powerful country in the world. With a fantastic wealth of material resources and a high industrialization, the American cities reflect the advantages and disadvantages of this process. The fourth chapter is concerned with the City of Houston: its historical, social and economical development. It includes an urban analysis and its relation to the family, the community and the city. It includes the actual problem of the community with its strong and fast technological improvements and the impact that the community receives which has to be expressed with new spatial order and relationship in the city. The last chapter is devoted to the design and development of a Houston community. It shows a community that provides to the people the means of a better way of living in. a fast-moving and dynamic society