Exploring several nonconventional interactions of ice-binding proteins

Traditionally, ice-binding proteins (IBPs), also known as antifreeze proteins (AFPs), have been defined by two universal activities: ice recrystallization inhibition and thermal hysteresis. However, there remains the possibility IBPs have other complementary functions given the diversity found within this protein group. This thesis explores some of these in both natural and applied settings, in the hopes of furthering our understanding of this remarkable group of proteins. Plant IBPs could function as part of a defensive strategy against ice nucleators produced by certain pathogens. To assess this hypothesis, recombinant IBPs from perennial ryegrass and purple false brome were combined with the ice nucleation protein (INP) from the plant pathogen, Pseudomonas syringae. Strikingly, the plant proteins depressed the freezing point of the bacterial INP, while a fish AFP could not, nor did the INPs have any effect on IBP activity. Thus, the interaction between these two different proteins suggests a role in plant defensive strategies against pathogenic bacteria as another IBP function. In addition, the potential use of hyperactive insect IBPs in organ preservation was investigated. Current kidney preservation techniques involve storing the organ at 4 °C for a maximum of 24 h prior to transplantation. Extending this “safe” time would have profound effects on renal transplants, however, ischemic injury is prevalent when storage periods are prolonged. Experiments described here allowed subzero preservation for 72 h with the addition of a beetle IBP to CryoStasis® solution. Kidneys stored using the traditional technique for 24 h and the method developed here for 72 h showed similar levels of biomarker enzymes, underscoring the potential utility of insect IBPs for future transplant purposes. Finally, IBP function in the freeze-tolerant gall fly, Eurosta solidaginis, was examined. Larvae representing the mid-autumn stage displayed ice-binding activity, suggesting an IBP is being expressed, possibly as a protective measure against freezing damage when fall temperatures can unpredictably drop. IBP activity was also observed in the larvae’s host plant, Solidago spp. Mass spectrometry analysis of ice-affinity purified plant extracts provided three candidate pathogenesis-related proteins that could be responsible for the detected activity, further demonstrating additional functions of IBPs.

BARRIERS AND SOLUTIONS TO OPEN SOURCE APPROPRIATE TECHNOLOGY FOR SUSTAINABLE DEVELOPMENT: INNOVATION THROUGH COLLABORATION

It has been proposed that the field of appropriate technology (AT) - small-scale, energy efficient and low-cost solutions, can be of tremendous assistance in many of the sustainable development challenges, such as food and water security, health, shelter, education and work opportunities. Unfortunately, there has not yet been a significant uptake of AT by organizations, researchers, policy makers or the mainstream public working in the many areas of the development sector. Some of the biggest barriers to higher AT engagement include: 1) AT perceived as inferior or ‘poor persons technology’, 2) questions of technological robustness, design, fit and transferability, 3) funding, 4) institutional support, as well as 5) general barriers associated with tackling rural poverty. With the rise of information and communication technologies (ICTs) for online networking and knowledge sharing, the possibilities to tap into the collaborative open-access and open-source AT are growing, and so is the prospect for collective poverty reducing strategies, enhancement of entrepreneurship, communications, education and a diffusion of life-changing technologies. In short, the same collaborative philosophy employed in the success of open source software can be applied to hardware design of technologies to improve sustainable development efforts worldwide. To analyze current barriers to open source appropriate technology (OSAT) and explore opportunities to overcome such obstacles, a series of interviews with researchers and organizations working in the field of AT were conducted. The results of the interviews confirmed the majority of literature identified barriers, but also revealed that the most pressing problem for organizations and researchers currently working in the field of AT is the need for much better communication and collaboration to share the knowledge and resources and work in partnership. In addition, interviews showcased general receptiveness to the principles of collaborative innovation and open source on the ground level. A much greater focus on networking, collaboration, demand-led innovation, community participation, and the inclusion of educational institutions through student involvement can be of significant help to build the necessary knowledge base, networks and the critical mass exposure for the growth of appropriate technology.