This research will: 1) Behaviourally assess the extent to which macaques can learn different forms of nonadjacent relationships in sequences generated by artificial grammars. 2) Use comparative neuroimaging to identify the brain areas involved in complex sequence processing in both macaques and humans and evaluate the correspondences and potential divergences across the species in key frontal, temporal and parietal brain areas. 3) Obtain neuroimaging connectivity data to evaluate the involvement of dorsal and ventral pathways involved in these processes across the species. To achieve these objectives I will: 1) test macaques and humans abilities to learn a suite of sequence processing tasks designed to independently increase sequencing complexity and working memory demands. 2) fMRI will be used to assess whether the neurobiological substrates involved in evaluating nonadjacent sequencing relationships are supported by similar or different brain areas across the species. 3) The fMRI re sults will be used to inform connectivity analyses to understand the pathways connecting these frontal and temporal lobe regions. This project has the potential to clarify how the frontal cortex supports cognitive functions involved in evaluating complex sequencing processing, a potential human language precursor process, and the extent to which macaques could serve to model these processes.

Prof Argent and co-workers have a long-standing history of investigating electrolyte secretion and water transport in isolated pancreatic ducts. The current proposal is designed to examine the regulation of the activities of anion exchangers on human pancreatic duct cells. First of all we would like to establish a polarized human pancreatic duct cell model on which we would perform our experiments. The best candidate for this model is the CFPAC-1 cells. These cells were isolated from a cystic fibrosis patient with the ?F508 mutation who developed pancreatic cancer. In addition, I will use CFPAC-1 cells stably transfected with the wild type CFTR or an empty vector. We are especially interested in the basolateral SLC4 and the apical SLC26 families of anion exchangers. I will use molecular biological techniques to find out whether the mRNAs and proteins of SLC4 (particularly SLC4A2) and SLC26 (particularly SLC26A3 and SLC26A6) family members are present in the ducts cells. Moreover, I will test whether these exchangers are indeed localized at the luminal and/or basolateral membranes using confocal immunocytochemistry. The functional expression of these anion exchangers will be established from intracellular pH changes. I will also check whether the expression or the stimulation of CFTR has an effect on the anion exchangers. Computer modelling studies indicate that the activity of the apical anion exchanger in the pancreatic duct cells are down-regulated once the luminal bicarbonate concentration reaches 80mM. I would like to test my hypothesis that the rise in luminal pH indeed down-regulates the activity of the apical exchanger. Concerning some of the difficulties of measuring the apical anion exchange, I have designed two experimental protocols to achieve this aim. Activity of the basolateral SLC4A2 anion exchanger must also be inhibited during HCO3- secretion. This prediction will be tested using a radiolabelled SO42- flux technique. By using our cellular model system it will be possible to establish whether a functional CFTR is required for the inhibitory effect on basolateral anion exchange activity. In conclusion, in the present project we will reveal important information about the regulation and activities of the SLC4 and SLC26 families of anion exchangers in human pancreatic duct cells. We expect that the results of this research will give insight to the complex mechanisms of bicarbonate secretion by the pancreatic ductal epithelium.

Bacterial plasmids encode centromere-like functions (called par loci) that actively segregate plasmid molecules to daughter cells before cell division. All plasmid-encoded par loci can be divided into several types that encode unrelated ATPases: Type I loci encode P-loop ATPases while the common Type II loci encode actin-like ATPases. Type II loci segregate plasmids by a mitotic-like process in which long actin filaments push plasmids apart. It is less well understood how the common Type I loci function. Type I ParA ATPases are related to MinD and, remarkably, oscillate in spiral-shaped structures. The oscillating ATPases position plasmids regularly within the cell. Moreover, the ParB/parC partitioning complex pairs plasmids in vitro. We will analyse the molecular mechanism by which par2 of plasmid pB171 segregates plasmids using a 3-stranded strategy: (i) with living cells we will use double, triple and quadruple labelling of plasmid, ParA, ParB and the bacterial nucleoid thus to gain information of how the components interact to move and position plasmids; (ii) using purified components, we will reconstitute the parABC system in vitro; (iii) finally, we will make attempts to solve the structure of not only the ParB/parC complex but also the super-structure of the ParA/ParB/parC complex.