Prevention of infectious diseases by vaccination is often limited because of the lack of safe, effective, and accessible vaccines. Traditional vaccines are expensive and require special conditions for storage, distribution, and administration. Plants have potential for large-scale production of a variety of inexpensive and highly effective recombinant proteins for biomedical and pharmaceutical applications, including subunit vaccines. There are several approaches for the production of vaccine antigens in plants, including transient expression systems based on Agrobacterium delivery of binary vectors or plant viral vectors, stable transgenic plants, and plant cell or tissue cultures. Axenic plant cultures maintained under defined physical and chemical conditions appear to be an attractive production platform when target proteins need to be synthesized in a fully controlled environment. Hairy root cultures meet the criteria for such a system. Hairy root cultures, generated from edible plants and producing target antigens, provide a potential approach for the development of vaccines for oral delivery. With this approach, there are no protein extraction and purification costs and the active biomolecule is protected by the plant cell wall during passage through the upper gastrointestinal tract. This allows for gradual release of antigen at mucosal surfaces in the gut. Lyophilized hairy root cultures expressing vaccine antigens can be stored at ambient temperature for extended periods of time, which should facilitate storage and distribution, ultimately allowing for large populations to be vaccinated.
Substrates that are cleaved to yield fluorescent products can be used to quickly quantify enzyme activity in vivo with image analysis or in vitro with fluorometry. This study was carried out to determine whether enzyme activity in rotifers is useful for assessing the physiological condition of rotifers. Neonates of Brachionus plicatilis Müller hatched from cysts were exposed to a concentration series of unionized ammonia ranging from 0 to 9.8 p.p.m., increasing seawater viscosity relative to 1.17 to control sea water by the addition of methyl cellulose and the addition of the protozoan Euplotes sp. to a density of 40 mL−1. Rotifer glucosidase and esterase activities decreased with increasing unionized ammonia and viscosity respectively. Activities of glucosidase and phospholipase decreased with increasing protozoan contamination. There was a significant relationship between enzyme activities and rotifer population growth. In vivo activities of certain rotifer enzymes can therefore serve as biomarkers for the rapid assessment of environmental stressors in rotifer mass cultures.
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