The identification and characterisation of Rab4 interacting proteins

Rab4 is a member of the Rab superfamily of small GTPases. It is localized to the early sorting endosome and plays a role in regulating the transport from this compartment to the recycling and degradative pathways. In order to further our understanding of the role Rab4 plays in endocytosis, a yeast two-hybrid screen was performed to identify putative Rab4 effectors. A constitutively active mutant of Rab4, Rab4Q67L, when used as bait to screen a HeLa cDNA library, identified a novel 80kDa protein that interacted with Rab4-GTP. This protein was called Rab Coupling Protein (RCP). RCP interacts preferentially with the GTP-bound form of Rab4. Subsequent work demonstrated that RCP also interacts with Rab11, and that this interaction is not nucleotide-depenedent. RCP is predominantly membrane-bound and localised to the perinuclear recycling compartment. Expression of a truncation mutant of RCP, that contains the Rab binding domain, in HeLa cells, results in the formation of an extensive tubular network that can be labelled with transferrin. These tubules are derived from the recycling compartment since they are inaccessible to transferrin when the ligand is internalised at 18oC. The truncation mutant-induced morphology can be rescued by overexpression of active Rab11, but not active Rab4. This suggests that RCP functions between Rab4 and Rab11 in the receptor recycling pathway, and may act as a ‘molecular bridge’ between these two sequentially acting small GTPases. Quantitative assays demonstrated that overexpression of the truncation mutant results in a dramatic inhibition in the rate of receptor recycling. Database analysis revealed that RCP belongs to a family of Rab interacting proteins, each characterised by a carboxy-terminal coiled-coil domain and an amino-terminal phospholipid-binding domain. KIAA0941, an RCP homologue, interacts with Rab11, but not with Rab4. Overexpression of its Rab binding domain also results in a tubular network, however, this tubulation cannot be rescued by active Rab11.

Applications of finite element computer modelling and thermal infrared remote sensing to the study of geothermal anomalies

Buried heat sources can be investigated by examining thermal infrared images and comparing these with the results of theoretical models which predict the thermal anomaly a given heat source may generate. Key factors influencing surface temperature include the geometry and temperature of the heat source, the surface meteorological environment, and the thermal conductivity and anisotropy of the rock. In general, a geothermal heat flux of greater than 2% of solar insolation is required to produce a detectable thermal anomaly in a thermal infrared image. A heat source of, for example, 2-300K greater than the average surface temperature must be a t depth shallower than 50m for the detection of the anomaly in a thermal infrared image, for typical terrestrial conditions. Atmospheric factors are of critical importance. While the mean atmospheric temperature has little significance, the convection is a dominant factor, and can act to swamp the thermal signature entirely. Given a steady state heat source that produces a detectable thermal anomaly, it is possible to loosely constrain the physical properties of the heat source and surrounding rock, using the surface thermal anomaly as a basis. The success of this technique is highly dependent on the degree to which the physical properties of the host rock are known. Important parameters include the surface thermal properties and thermal conductivity of the rock. Modelling of transient thermal situations was carried out, to assess the effect of time dependant thermal fluxes. One-dimensional finite element models can be readily and accurately applied to the investigation of diurnal heat flow, as with thermal inertia models. Diurnal thermal models of environments on Earth, the Moon and Mars were carried out using finite elements and found to be consistent with published measurements. The heat flow from an injection of hot lava into a near surface lava tube was considered. While this approach was useful for study, and long term monitoring in inhospitable areas, it was found to have little hazard warning utility, as the time taken for the thermal energy to propagate to the surface in dry rock (several months) in very long. The resolution of the thermal infrared imaging system is an important factor. Presently available satellite based systems such as Landsat (resolution of 120m) are inadequate for detailed study of geothermal anomalies. Airborne systems, such as TIMS (variable resolution of 3-6m) are much more useful for discriminating small buried heat sources. Planned improvements in the resolution of satellite based systems will broaden the potential for application of the techniques developed in this thesis. It is important to note, however, that adequate spatial resolution is a necessary but not sufficient condition for successful application of these techniques.

Creating religious place in Ireland: Hindu public places of worship and the Indian sculpture park

This paper examines the creation of religious place. It argues that the designation of a place as “religious” is a subjective and creative act which is dependent upon the perception and past, or memory, of the viewer. The paper focuses specifically on the creation of public places of worship by Hindu groups in the Dublin city area of Ireland and on the varied perceptions of the Indian Sculpture Park in County Wicklow. The creation of public places of worship results in places classified as “religious” due to the intention of the creator, the terminology used and the types of activities that take place in the space. This is in contrast to places such as the Indian Sculpture Park in County Wicklow which was created as a secular space but which is viewed by some Hindus as an outdoor temple due to the presence of sculptures of the Hindu deity Ganesh. Other Hindus do not view the space as having any religious significance and so its religiosity is contested. This points to the fact that the creation of religious place is a creative act of interpretation which is dependent upon the perception and past of the viewer and which changes over time.

System packaging & integration for a swallowable capsule using a direct access sensor

Technological developments in biomedical microsystems are opening up new opportunities to improve healthcare procedures. Swallowable diagnostic capsules are an example of this. In this paper, a diagnostic capsule technology is described based on direct-access sensing of the Gastro Intestinal (GI) fluids throughout the GI tract. The objective of this paper is two-fold: i) develop a packaging method for a direct access sensor, ii) develop an encapsulation method to protect the system electronics. The integrity of the interconnection after sensor packaging and encapsulation is correlated to its reliability and thus of importance. The zero level packaging of the sensor was achieved by using a so called Flip Chip Over Hole (FCOH) method. This allowed the fluidic sensing media to interface with the sensor, while the rest of the chip including the electrical connections can be insulated effectively. Initial tests using Anisotropic Conductive Adhesive (ACA) interconnect for the FCOH demonstrated good electrical connections and functionality of the sensor chip. Also a preliminary encapsulation trial of the flip chipped sensor on a flexible test substrate has been carried out and showed that silicone encapsulation of the system is a viable option.

Wireless sensor node design for heterogeneous networks

Two complementary wireless sensor nodes for building two-tiered heterogeneous networks are presented. A larger node with a 25 mm by 25 mm size acts as the backbone of the network, and can handle complex data processing. A smaller, cheaper node with a 10 mm by 10 mm size can perform simpler sensor-interfacing tasks. The 25mm node is based on previous work that has been done in the Tyndall National Institute that created a modular wireless sensor node. In this work, a new 25mm module is developed operating in the 433/868 MHz frequency bands, with a range of 3.8 km. The 10mm node is highly miniaturised, while retaining a high level of modularity. It has been designed to support very energy efficient operation for applications with low duty cycles, with a sleep current of 3.3 μA. Both nodes use commercially available components and have low manufacturing costs to allow the construction of large networks. In addition, interface boards for communicating with nodes have been developed for both the 25mm and 10mm nodes. These interface boards provide a USB connection, and support recharging of a Li-ion battery from the USB power supply. This paper discusses the design goals, the design methods, and the resulting implementation.

Effect of solder volume on joint shape with variable chip-to-board contact pad ratio

The objective of this paper is to investigate the effect of the pad size ratio between the chip and board end of a solder joint on the shape of that solder joint in combination with the solder volume available. The shape of the solder joint is correlated to its reliability and thus of importance. For low density chip bond pad applications Flip Chip (FC) manufacturing costs can be kept down by using larger size board pads suitable for solder application. By using “Surface Evolver” software package the solder joint shapes associated with different size/shape solder preforms and chip/board pad ratios are predicted. In this case a so called Flip-Chip Over Hole (FCOH) assembly format has been used. Assembly trials involved the deposition of lead-free 99.3Sn0.7Cu solder on the board side, followed by reflow, an underfill process and back die encapsulation. During the assembly work pad off-sets occurred that have been taken into account for the Surface Evolver solder joint shape prediction and accurately matched the real assembly. Overall, good correlation was found between the simulated solder joint shape and the actual fabricated solder joint shapes. Solder preforms were found to exhibit better control over the solder volume. Reflow simulation of commercially available solder preform volumes suggests that for a fixed stand-off height and chip-board pad ratio, the solder volume value and the surface tension determines the shape of the joint.

Miniaturised multi-MEMS sensor development

Complex systems, from environmental behaviour to electronics reliability, can now be monitored with Wireless Sensor Networks (WSN), where multiple environmental sensors are deployed in remote locations. This ensures aggregation and reading of data, at lower cost and lower power consumption. Because miniaturisation of the sensing system is hampered by the fact that discrete sensors and electronics consume board area, the development of MEMS sensors offers a promising solution. At Tyndall, the fabrication flow of multiple sensors has been made compatible with CMOS circuitry to further reduce size and cost. An ideal platform on which to host these MEMS environmental sensors is the Tyndall modular wireless mote. This paper describes the development and test of the latest sensors incorporating temperature, humidity, corrosion, and gas. It demonstrates their deployment on the Tyndall platform, allowing real-time readings, data aggregation and cross-correlation capabilities. It also presents the design of the next generation sensing platform using the novel 10mm wireless cube developed by Tyndall.