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27. Progress of vitrification: From single cell to multiple objects

Cryobiology 2010年第3期61卷 370-370页

作者： L.L. Kuleshova S.S. Gouk R. Magalhaes F. Wen Y.N. Wu G.S. Dawe H. Yu Low Temperature Preservation Unit National University Medical Institutes Yong Loo Lin School of Medicine National University of Singapore Singapore Department of Orthopaedic Surgery NUS Tissue Engineering Program Yong Loo Lin School of Medicine National University of Singapore Singapore Department of Pharmacology Yong Loo Lin School of Medicine National University of Singapore Singapore Neurobiology and Ageing Centre for Life Sciences National University of Singapore Singapore Department of Physiology Yong Loo Lin School of Medicine National University of Singapore Singapore Institute of Bioengineering and Nanotechnology Agency for Science Technology and Research Singapore

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GFP synthesis in giant liposomes using the in vitro translation system of Thermococcus kodaks

GFP synthesis in giant liposomes using the in vitro translat...

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20th Anniversary MHS 2009 and Micro-Nano Global COE - 2009 International Symposium on Micro-NanoMechatronics and Human science

作者： Yamaji, Kazuaki Kanai, Tamotsu Nomura, Shin-Ichiro M. Akiyoshi, Kazunari Negishi, Makiko Atomi, Haruyuki Yoshikawa, Kenichi Imanaka, Tadayuki Graduate School of Engineering Kyoto University Nishikyo-ku Kyoto 615-8510 Japan Institute for Integrated Cell-Material Sciences Kyoto University Sakyo Kyoto 606-8501 Japan Institute of Biomaterials and Bioengineering Tokyo Medical and Dental University Chiyoda-ku Tokyo 101-0062 Japan Graduate School of Science Kyoto University Sakyo-ku Kyoto 606-8502 Japan College of Life Sciences Ritsumeikan University Kusatsu Shiga 525-8577 Japan

ISBN: (纸本)9781424450954

The possibilities of an in vitro translation system, based on cell components of the hyperthermophilic archaeon, Thermococcus kodakaraensis, to utilize at a lower temperature range were examined. Green fluorescence protein (GFP) was used as the reporter protein. Through optimization of several experimental conditions, the yield of active GFP production at 40°C increased 5-fold. The optimized system was encapsulated in liposome made by passing through the oil/water interfaces. The prepared liposomes were incubated at 40°C for 90 min, and fluorescence was examined by laser confocal microscopy. A significant increase in average fluorescence intensity was observed in liposomes with GFP mRNA, indicating that the T. kodakaraensis in vitro translation system is applicable for protein production within giant liposomes. These artificial cell models should provide the means to reconstitute various cell functions from a synthetic biology approach. ©2009 IEEE.

关键词： Liposomes

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Macromol. Rapid Commun. 13/2010

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Macromolecular Rapid Communications 2010年第13期31卷

作者： Mai Sanjoh Shigehiro Hiki Yan Lee Makoto Oba Kanjiro Miyata Takehiko Ishii Kazunori Kataoka Department of Bioengineering Graduate School of Engineering The University of Tokyo Japan Department of Materials Engineering Graduate School of Engineering The University of Tokyo 7‐3‐1 Hongo Bunkyo‐ku Tokyo 113‐8656 Japan Core Research Program for Evolutional Science and Technology (CREST) Japan Science and Technology Agency (JST) Tokyo Japan Department of Chemistry Seoul National University Sillim‐dong Gwanak‐gu Seoul 151‐742 Korea Department of Clinical Vascular Regeneration Graduate School of Medicine The University of Tokyo 7‐3‐1 Hongo Bunkyo‐ku Tokyo 113‐8655 Japan Center for Disease Biology and Integrative Medicine Graduate School of Medicine The University of Tokyo 7‐3‐1 Hongo Bunkyo‐ku Tokyo 113‐0033 Japan Center for NanoBio Integration The University of Tokyo 7‐3‐1 Hongo Bunkyo‐ku Tokyo 113‐8656 Japan

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Tandem multimer expression of angiotensin I-converting enzyme inhibitory peptide in Escherichia coli

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Biotechnology Journal 2009年第9期4卷 1345-1356页

作者： Fida, Hasan M. Kumada, Yoichi Terashima, Masaaki Katsuda, Tomohisa Katoh, Shigeo Department of Molecular Science and Material Engineering Graduate School of Science and Technology Faculty of Engineering Kobe Japan Department of Chemistry and Materials Technology Kyoto Institute of Technology Kyoto Japan Department of Biosphere Sciences School of Human Science Kobe College Nishinomiya Japan Department of Chemical Engineering Hanyang University 1271 Sa-3-dong Sangrok-gu Ansan 426-791 Korea Republic of Department of Process Engineering and Applied Science Food Science and Technology Program Dalhousie University 1360 Barrington Street Halifax NS B3J 2X4 Canada

It is common for small tandem peptide multimer genes to be indirectly inserted into expression vectors and fused with a protein tag. In this study, a multimer of the tandem angiotensin I-converting enzyme inhibitory peptide (ACE-IP) gene was directly transferred to a commercially available vector and the designed gene was expressed as a repeated peptide in Escherichia coli BL21(DE3)pLysS. The process further developed in our study was the construction of six-repeated ACE-IP synthetic genes and their direct insertion. Protein expression in inclusion bodies was confirmed by SDS-PAGE and Western blot. Acid hydrolysis of inclusion bodies produced single-unit peptides through cleavage of the aspartyl-prolyl bonds. This cleaved recombinant peptide (rACE-IP) was purified using immuno-affinity chromatography followed by reversed phase-HPLC. 105-115 mg of the lyophilized recombinant peptide was obtained from 1 L E. coli culture. In vitro biological activity of rACE-IP was indistinguishable from that of the natural peptide produced by hydrolysis in artificial gastric juice or by acidic hydrolysis. The rACE-IP prepared by recombinant DNA technology and solid-phase synthesis methods showed a similar IC50. This strategy could be used for the expression of important peptides, which have N-terminal proline (P) and C-terminal aspartic acid residues (D) for commercial applications, e.g. functional foods and drinks. © 2009 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim.

关键词： Peptides

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GFP synthesis in giant liposomes using the in vitro translation system of Thermococcus kodakaraensis

GFP synthesis in giant liposomes using the in vitro translat...

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International Symposium on Micromechatronics and Human science (MHS)

作者： Kazuaki Yamaji Tamotsu Kanai Shin-ichiro M. Nomura Kazunari Akiyoshi Makiko Negishi Haruyuki Atomi Kenichi Yoshikawa Tadayuki Imanaka Graduate School of Engineering Kyoto University Nishikyo Kyoto Japan Institute for Integrated Cell-Material Sciences Kyoto University Sakyo Kyoto Japan Institute of Biomaterials & Bioengineering Tokyo Medical and Dental University Chiyoda Tokyo Japan Graduate School of Science Kyoto University Sakyo Kyoto Japan College of Life Sciences Ritsumeikan University Kusatsu Shiga Japan

关键词： In vitro Fluorescence Proteins Cells (biology) Hyperthermia Temperature distribution Production Petroleum Laser modes Microscopy

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Noninvasive Drug Delivery Using Ultrasound: Targeting Melanoma Using siRNA Against Mutant (V600E) B‐Raf

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AIP Conference Proceedings 2009年第1期1113卷 423-427页

作者： Melissa A. Tran Raghavendra Gowda Eun‐Joo Park James Adair Nadine Smith Mark Kester Gavin P. Robertson aDepartment of Pharmacology The Pennsylvania State University College of Medicine Hershey PA 17033 fDepartment of Bioengineering Pennsylvania State University University Park PA USA dThe Penn State Melanoma Therapeutics Program Hershey PA 17033 gDepartment of Material Science and Engineering Pennsylvania State University University Park PA USA bDepartment of Pathology The Pennsylvania State University College of Medicine Hershey PA 17033 cDepartment of Dermatology The Pennsylvania State University College of Medicine Hershey PA 17033 eThe Foreman Foundation for Melanoma Research Hershey PA 17033

Melanoma is the most deadly form of skin cancer. Currently early surgical removal is the best treatment option for melanoma patients with little hope of successful treatment of late stage melanoma. Clearly new treatment options must be explored. Topical administration of drugs provides the advantage of being able to apply large quantities of drug in close proximity to the tumor without the issue of systemic side effects. However, the natural barrier formed by the skin must first be overcome for topical treatment to become a viable option. With this in mind we have sought to use low‐frequency ultrasound to transiently permeabilize the stratum corneum and successfully deliver liposomal siRNA to melanoma cells residing at the basement membrane. B‐Raf is one of the most frequently activated genes in melanoma, making it an ideal candidate for targeting via siRNA. The novel liposomes used in this study load siRNA, protect if from the outside environment and lead to knockdown of target message. Combining ultrasound with liposomal siRNA we show that siRNA can be delivered into melanoma cells. Additionally, we show that siRNA to mutant B‐Raf can effectively inhibit melanoma growth in reconstructs and in mice by 60% and 30% respectively. Therefore, ultrasound with liposomal siRNA is a potentially valuable treatment option for melanoma patients.

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Microchip analysis of cellular respiratory activity on PDMS membrane having gas permselective property

Microchip analysis of cellular respiratory activity on PDMS ...

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12th International Conference on Miniaturized Systems for Chemistry and Life sciences, MicroTAS 2008

作者： Shirai, T. Sakata, T. Takai, M. Miyahara, Y. Ishihara, K. Department of Material Engineering School of Engineering University of Tokyo Japan Biomaterial Center National Institute for Material Science Tsukuba Japan Department of Bioengineering School of Engineering University of Tokyo Japan

A new microchip device for real-time analysis of cellular respiratory activity on poly(dimethylsiloxane) (PDMS) membrane with integration of carbon dioxide sensor based on pH-sensitive field-effect transistor (pH-FET) was developed. Controlling the thickness of PDMS membrane, response curves of the CO2 gas under soda water flow and respiratory activity of HeLa cells on PDMS substrate were monitored as a pH-shift. Our results suggest that the pH-FET integrated microdevice having gas permselective PDMS membrane can contribute critically to basic understanding of biological mechanisms in the field of tissue engineering. © 2008 CBMS.

关键词： Carbon dioxide

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GEO: A web-based event registration and communication tool

GEO: A web-based event registration and communication tool

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2008 ASEE Annual Conference and Exposition

作者： Bowman, David Stephan, Elizabeth Benson, Lisa Clemson University General Engineering Program Department of Engineering and Science Education Clemson University School of Computing Clemson University Department of Engineering and Science Education Department of Bioengineering Clemson University

The first-year engineering courses at Clemson University involve various out-of-class requirements, such as departmental tours, exams, team projects, and presentations. Enrollment in the first semester course typically exceeds 850 students, and the number of students who can attend any single out-of-class event has physical and logistical limitations. Additionally, because these students are unfamiliar with course guidelines, procedures, and each other, straightforward communication tools are vital to their success. A web-based system, GEO (General engineering Online), was developed using PHP1 (a programming language used to dynamically generate web pages) and MySQL (a relational database management system). The system allows pre-registration for required out-of-class activities (department tours, advisor appointments, and team project work sessions). It also provides a portal for communication between students, instructors and advisors (peer evaluations, surveys, email confirmations of appointments, and student/team management). This study examines the capabilities of the GEO system from the viewpoint of students and faculty, as well as its pedagogical implications. The program and data structure are described, along with sample statistics generated by the system. Future directions include the development of GEOPort, an individual and team work documentation tool. This will transform GEO from a registration and communication tool into an education research tool. Team members will be able to post text, pictures, or video in response to instructor-posted materials, and to document their progress on projects and assignments. Rubrics can be developed to grade online submissions, and to track student approaches to design, problem-solving, and team dynamics. © American Society for engineering Education, 2008.

关键词： Students

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A unique research experience in bioengineering education for undergraduates in the VaNTH REU

A unique research experience in bioengineering education for...

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2008 ASEE Annual Conference and Exposition

作者： Linsenmeier, Robert Alley, Jean Hirsch, Penny Klein, Stacy Greenberg, Julie Bourgeois, Mark Northwestern University Robert R. McCormick School of Engineering and Applied Science Weinberg College of Arts and Sciences VaNTH Engineering Research Center in Bioengineering Educational Technologies Vanderbilt University Writing Program at Northwestern Outreach Vanderbilt University School of Engineering Department of Biomedical Engineering Radiological Sciences and Teaching and Learning Massachusetts Institute of Technology Philosophy Loyola University Chicago

Most Research Experience for Undergraduates (REU) programs are designed to provide summer research opportunities in a particular domain of engineering or science. The VaNTH REU has been unique in focusing instead on education research projects in bioengineering. These projects allow students from various fields to gain a different perspective on their education while making the decisions about content and pedagogy that instructors usually make. After an introductory session at Vanderbilt, students spend the next nine weeks working with mentors from Vanderbilt University, Northwestern University, Massachusetts Institute of Technology or the University of Texas at Austin. While at these sites, students meet weekly by video- or tele-conference to report on their projects. Work on ethics and communication were integrated into the REU experience. The REU students, who are mostly in engineering, have become engaged in the process and methods of engineering education research and, in many cases, have made substantial contributions to the development and/or classroom evaluation of educational materials. At the same time, they have learned a particular field of bioengineering more deeply. This paper discusses the VaNTH REU program and illustrates the contributions of REU students to successful innovations in bioengineering pedagogy. While it would be difficult to replicate the VaNTH REU program in its entirety, many of its components are transferable and could help students who are considering faculty careers or graduate school in engineering education. © American Society for engineering Education, 2008.

关键词： Students

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Poly(ethylene glycol)-Protected, pH-dependent liposomes enhance RNAi delivery

Poly(ethylene glycol)-Protected, pH-dependent liposomes enha...

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2008 AIChE Annual Meeting, AIChE 100

作者： Auguste, Debra T. Furman, Kay Wong, Andrew D. Fuller, Jason Armes, Steven P. Deming, Timothy Langer, Robert S. School of Engineering and Applied Sciences Harvard University 29 Oxford St. Cambridge MA 02138 United States Department of Material Science and Engineering Massachusetts Institute of Technology 77 Massachusetts Avenue Cambridge MA 02139 United States Department of Chemical Engineering Massachusetts Institute of Technology 45 Carleton St. Cambridge MA 02142 United States Department of Chemistry Dainton Building University of Sheffield Sheffield United Kingdom Department of Bioengineering UCLA Bolter Hall Los Angeles CA 90095 United States

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