Search

Mammalian cloning by somatic nuclear transfer has great potential for developing medical applications such as biopharmaceuticals and generation of tissues for transplantation. For agricultural applications, it allows the rapid dissemination of genetic gain in livestock breeding. The maximisation of that potential requires improvements to overall cloning technology, especially with respect to increasing cloning efficiency and throughput rates in cloned embryo production. A zona-free embryo reconstruction system was developed to increase cloning throughput and ease of operation. Central to this system is a modified electrofusion procedure for nuclear transfer. Cytoplast-donor cell couplets were placed in a custom-designed 'parallel plate' electrode chamber. A 1 MHz sinusoidal AC dielectrophoresis alignment electric field of 6-10 kV m(-1) was applied for 5-10s. The couplets were then fused using 2 x 10 micros rectangular DC-field pulses (150-200 kV m(-1)), followed by application of the AC field (6-10 kV m(-1)) for another 5-10 s. Fusion was performed in hypoosmolar buffer (210 mOsm). Automated alignment of up to 20 couplets at a time has been achieved, resulting in greatly improved fusion throughput rates (2.5-fold increase) and improved fusion yields (1.3-fold increase), compared with commonly followed zona-intact protocols.

The avoidance of house dust mite allergens is a major area of interest and essentially requires a significant removal of these allergens from the immediately respirable air. Electrostatic attraction and anchoring of particulate matter using electret polymers is commonly used for air filtration purposes. This effect is investigated for its possible use in domestic allergen avoidance. Polypropylene electret, heat-treated electret and non-electret, and wool and nylon fibre samples were soiled with house dust known to contain Der p 1 allergen. These samples were vacuumed at three air face velocities. The proportions of released and anchored dust were calculated. Released dust was collected and analysed for Der p 1 concentration and compared to stock dust values. Results showed that compared to uncharged fibres at least 95% more dust remained anchored in the electret fibres. Also, overall Der p 1 release was reduced by more than 49%. Der p 1 allergen concentrations in the collected dust were relatively constant for all the fibres tested, indicating no selective attraction or repulsion of Der p 1 allergen carrying particles in the experimental dust. The consistently high dust anchoring ability of the electret fibres could be used in many domestic products that are known to harbour particulate allergens, to reduce their release and inhalation.

Control of the house dust mite allergen has received considerable attention owing to its importance in some allergic diseases. One aspect of dust mites and their allergen-carrying faecal particles that has not been reported on, which may have allergen control applications, is the electrostatic charge they carry in the natural environment. To promote tribo-electric charging, household dust containing dust mite allergen and live house dust mites are separately agitated while in contact with either polypropylene, nylon or earthed metal. The charged dust and mites are subsequently subjected to electrostatic separation and collection. Results for concentrations of the house dust mite allergen, Der p1, indicate that, when subjected to nylon, Der p1 carrier particles appear to be predominantly positively charged. Similarly, when subjected to polypropylene, Der p1 carrier particles also appear to be positively charged. Reduction of excess free charge by agitation against earthed metal does not appear to affect the observed charging characteristics, indicating that the positive charge may be bound or inherent in the Der p1 carrier particles. In contrast, house dust mites exposed to nylon appear to be generally charging negative, whereas mites exposed to polypropylene appear to be charging positive. The observed electrostatic characteristics of the mites and Der p1 carrying particles will be useful in the future development of electrostatic allergen control methods.