Microheated substrates for patterning cells and controlling development

Here, we seek to control cellular development by devising a means through which cells can be subjected to a microheated environment in standard culture conditions. Numerous techniques have been devised for controlling cellular function and development via manipulation of surface environmental cues at the micro- and nanoscale. It is well understood that temperature plays a significant role in the rate of cellular activities, migratory behavior (thermotaxis), and in some cases, protein expression. Yet, the effects and possible utilization of micrometer-scale temperature fields in cell cultures have not been explored. Toward this end, two types of thermally isolated microheated substrates were designed and fabricated, one with standard backside etching beneath a dielectric film and another with a combination of surface and bulk micromachining and backside etching. The substrates were characterized with infrared microscopy, finite element modeling, scanning electron microscopy, stylus profilometry, and electrothermal calibrations. Neuron culture studies were conducted on these substrates to 1) examine the feasibility of using a microheated environment to achieve patterned cell growth and 2) selectively accelerate neural development on regions less than 100$mu m$wide. Results show that attached neurons, grown on microheated regions set at 37$~^circ C$, extended processes substantially faster than those incubated at 25$~^circ C$on the same substrate. Further, unattached neurons were positioned precisely along the length of the heater filament (operating at 45$~^circ C$) using free convection currents. These preliminary findings indicate that microheated substrates may be used to direct cellular development spatially in a practical manner.$hfillhbox[1414]

Biotechnology approaches to overcome biotic and abiotic stress constraints in legumes

Biotic and abiotic stresses cause significant yield losses in legumes and can significantly affect their productivity. Biotechnology tools such as marker-assisted breeding, tissue culture, in vitro mutagenesis and genetic transformation can contribute to solve or reduce some of these constraints. However, only limited success has been achieved so far. The emergence of “omic” technologies and the establishment of model legume plants such as Medicago truncatula and Lotus japonicus are promising strategies for understanding the molecular genetic basis of stress resistance, which is an important bottleneck for molecular breeding. Understanding the mechanisms that regulate the expression of stress-related genes is a fundamental issue in plant biology and will be necessary for the genetic improvement of legumes. In this review, we describe the current status of biotechnology approaches in relation to biotic and abiotic stresses in legumes and how these useful tools could be used to improve resistance to important constraints affecting legume crops.E. Prats is funded by an European Marie Curie Reintegration Grant, N. Rispail by (FP5) Eufaba project. Our work in this area is supported by Spanish CICYT project AGL-2002-03248 and European Union project FP6-2002-FOOD-1-506223. K. Singh’s work in this area is supported in part by the Grains Research and Development Corporation (GRDC) and the Department of Education, Science and Training (DEST) in Australia.Peer reviewe