Platelet-derived growth factor D-chain (PDGF-D) is the newest member of the PDGF family of mitogens and chemo-attractants; it is expressed in a wide variety of cell types, including vascular smooth muscle cells (SMCs). The molecular mechanisms regulating PDGF-D transcription are unknown. Here I investigated the effects of angiotensin II (ATIl) and IL-1 beta on the transcription of PDGF-D and changes in vascular SMCs phenotype. Primer extension analysis mapped a single transcriptional start site to the ccAG CGC motif of PDGF-D promoter. Several potential transcription factor binding sites such as SpI, Ets-1, NF-??B, IRF-1, p53, Smad4 and AP1 were located in the proximal 1168bp of the PDGF-D promoter. ATII-inducible Ets-1 and PDGF-D gene expression is mediated via H202. IL-I beta supresses PDGF-D promoter activity, mRNA and protein expression in SMCs through NF-??B p65, IRF-1 and HDAC1, which form complex in the PDGF-D promoter. This study provides the first direct link between NF-KB and the PDGF-D promoter, IRF-1 with any member of the PDGF family and a new example of HDAC mediated inhibition of gene expression. In summary, this study investigates for the first time the mechanisms mediating the transcriptional regulation of PDGF-D in vascular SMCs. This provides valuable insights into the molecular control of vascular phenotype, and opens up potential opportunities for therapeutic intervention.

Mud crabs of the genus Scylla are widely exploited for aquaculture in the Asia- Pacific region. In the current study, a series of in vivo experiments were carried to assess the protein requirement, protein sparing effects of starch and the capacity of Scylla serrata to digest diets that contained different animal and plant-based feed meals and different levels and types of starch. Results from a protein requirement study indicated that juvenile S. serrata fed diets containing 45% or 55% protein demonstrated significantly higher growth responses than those fed the diet containing 25% protein. The subsequent study was carried out to determine if responses to dietary protein could be influenced by using purified wheat, potato, rice or corn starch to manipulate the gross energy level of fishmeal- based diets (18 or 15.5 MJ kg-1), i.e., to see if starch had a protein sparing effect in these animals. Overall, growth responses in this study appeared to be positively correlated with the level of protein in the diet with the highest growth rates achieved using diets containing 45% protein, regardless of the energy level of the diet. In addition, at a dietary protein level of 40% there was no evidence that the source of starch had any significant impact on growth performance or feed utilisation suggesting it had no protein sparing effect. By contrast, it was found that growth of juvenile S. serrata was strongly correlated with the intake of digestible dietary protein. The investigation of the capacity of sub-adult S. serrata to digest different animal and plant- based feed meals showed that apparent dry matter digestibility (ADMD) and apparent gross energy digestibility (AGED) values were not significant different for most selected feed meals (cotton seed, poultry, canola, fishmeal, soybean, and lupin meal). Apparent crude protein digestibility (ACPD) for all test feed meals were relatively high (86-96%). A subsequent study was carried out to determine if purified starch from different sources influenced the digestibility of fishmeal based diets. Overall, most diets containing starch were readily digested by mud crabs. In particular, there were no negative impacts on the digestibility of major nutrients (e.g. protein) observed following the inclusion of wheat, rice or corn starch in formulated feeds. Nevertheless, the apparent starch digestibility (ASD) of wheat starch decreased significantly as the inclusion level was increased from 15% to 60%, although there was no significant effect on ACPD values. At a 30% inclusion level, the ASD of diets containing different starches decreased in the order corn > wheat > potato = rice. Moreover, ACPD values were significantly higher for diets containing corn or rice starch than for those containing wheat or potato starch. The capacity of another species of mud crab commonly exploited for aquaculture in South East Asia, S. paramamosain, to digest the local plant-based ingredients (defatted soybean meal, rice bran, cassava and corn flour) was also conducted in Vietnam. Overall, the findings of this study showed that at a 30% inclusion level diets containing soybean meal or rice bran were well digested by mud crabs. In particular, the ACPD and AGED values for all diets containing soybean meal were not significantly different from the fishmeal based reference diet. Likewise, all digestibility values for the diet containing 30% rice bran were relatively high and not significantly different from the reference diet. By contrast, diets containing cassava flour appeared to be poorly utilised since their digestibility values for all parameters were lower than those from other testingredients. In summary, the apparent digestibility of dry matter, protein and energy was in the following order (from most to least digestible) soybean meal ~ rice bran > corn flour > cassava flour. In the next study the effects of attractants in diets (chicken meal, betaine, tuna oil and bait enhancer), temperature (26.5oC, 28.5oC and 30.5oC), sex (female and male) and size (small, medium and large) on feeding responses of S. serrata were investigated. Significant differences were observed in the behavioral responses of mud crabs to diets containing different attractants. Specifically, consumption of diets with chicken meal or betaine was significantly higher than for other treatments. With the exception of betaine, no significant difference in food consumption was observed when attractant inclusion levels were raised from 2% to 5%. Overall, small crabs consumed significantly more of the ration (as a percentage of body weight) than larger crabs. Temperature showed a significant impact on most behaviour of mud crabs, excepting continuation response and there was some evidence that females were significantly more active than males. Light intensity was considered as a main factor effect to crab response since there were extremely high percentage time of crab spent in half-shaded of the Y –maze which valued at 95.6%, 93.8 and 94.4% (corresponded to small, medium and large size respectively) in comparison to those of crabs spent in the unshaded side. Overall, the findings from these studies demonstrated that mud crabs have a high capacity to digest a range of plant based feed ingredients. In particular, soybean meal appeared to be well digested by both species of mud crabs examined. It was also shown that a range of purified starches were well digested by S. serrata although starch inclusion in diets did not appear to reduce the requirement for protein to promote growth. Subsequent attractant studies demonstrated that chicken meal and betaine produced significantly elevated feeding responses and food consumption when added to diets. Based on these results we propose that these ingredients can be utilised to increase the attractiveness and consumption of artificial mud crab feeds.