Decay or defeat ? : an inquiry into the Portuguese decline in Asia 1580-1645

Already in the 1590s the Portuguese in Asia looked upon the Dutch as a threat and most historiography has not been able to get away from the part that the Dutch played in the Indo-Portuguese drama. The decline of the Portuguese-Asian empire was however the result of endogenous and extrageneous developments, in Asia as well as in Brazil, Africa and Europe. For an analysis of these developments a multi-linear approach has been chosen in the form of, what one could call, a revolving stage. Each scene, or rather each chapter, produces in the end a different answer to the same question. The first five chapters discuss the social and financial fundamentals of the Portuguese 'empire' overseas and the position of the Portuguese in Asia, in terms of population, trade and military power. Special emphasis has been laid on the relationship of the so called New Christian Portuguese with the Castilian crown and their particular role in the trade with India. It will be shown that, to them, satisfying the need for silver of the Habsburg monarchy became a more attractive proposition than investment in the Carreira da India. This and other developments in Asia undermined the position of the Estado da India and of the private Portuguese traders in Asia, before the Dutch became a serious threat to them. The next three chapters are concerned with the Dutch: it will be shown that their active role in the Iberian and the Luso-Atlantic trade did not exclude an aggressive mood in Asia, or vice-versa. Dutch aggression in Asia was in the first instance prompted and legalized by the States-General, but commercial considerations also caused the bewindhebbers of the VOC to adopt a bellicose way of thinking and writing. However, it will be shown that apart from some acts of piracy and privateering against Portuguese ships and attacks on the Portuguese forts on the Moluccan islands, Dutch violence was mainly directed against the Spanish in the Philippines and the Chinese trade with these islands. On the other hand, whereas the conquest of the Moluccan spice trade became a first priority, the VOC was unable to prevent the Spaniards from taking over many Portugue-se forts and even had to accept that the Portuguese spice merchants moved to Macassar, where for a long time they stayed out of reach of the Company. In the discussion of the Dutch commercial and military initiatives many of the paradigms around the rise of the Dutch empire in Asia will be punctured. In the first forty years of its existence the VOC was far from the effective business organization or war machine that many writers have made it to be. Many of the glorified feats of arms can only be described as defeats or a waste of manpower, ships and money. Finally, the ninth chapter concentrates on the Asian environment in which the Luso-Dutch confrontation took place. During the period under review, major shifts in the local political situation were caused by the southward expansion of the Moghul empire, the rise of the Nayaks of Ikkeri in Kanara, the expansion of Persia under Shah Abbas, the unification and state formation in Japan under the Tokugawas and finally, the Manchu conquest of China. As far as the Portuguese were concerned, all these developments, each in their own way, worked in the same, negative, direction

Functional Interactions between KCNE1 C-Terminus and the KCNQ1 Channel

The KCNE1 gene product (minK protein) associates with the cardiac KvLQT1 potassium channel (encoded by KCNQ1) to create the cardiac slowly activating delayed rectifier, IKs. Mutations throughout both genes are linked to the hereditary cardiac arrhythmias in the Long QT Syndrome (LQTS). KCNE1 exerts its specific regulation of KCNQ1 activation via interactions between membrane-spanning segments of the two proteins. Less detailed attention has been focused on the role of the KCNE1 C-terminus in regulating channel behavior. We analyzed the effects of an LQT5 point mutation (D76N) and the truncation of the entire C-terminus (Δ70) on channel regulation, assembly and interaction. Both mutations significantly shifted voltage dependence of activation in the depolarizing direction and decreased IKs current density. They also accelerated rates of channel deactivation but notably, did not affect activation kinetics. Truncation of the C-terminus reduced the apparent affinity of KCNE1 for KCNQ1, resulting in impaired channel formation and presentation of KCNQ1/KCNE1 complexes to the surface. Complete saturation of KCNQ1 channels with KCNE1-Δ70 could be achieved by relative over-expression of the KCNE subunit. Rate-dependent facilitation of K+ conductance, a key property of IKs that enables action potential shortening at higher heart rates, was defective for both KCNE1 C-terminal mutations, and may contribute to the clinical phenotype of arrhythmias triggered by heart rate elevations during exercise in LQTS mutations. These results support several roles for KCNE1 C-terminus interaction with KCNQ1: regulation of channel assembly, open-state destabilization, and kinetics of channel deactivation

Heterologous Overexpression and Mutagenesis of the Human Bile Salt Export Pump (ABCB11) Using DREAM (Directed REcombination-Assisted Mutagenesis)

Homologous recombination in Saccharomyces cerevisiae is a well-studied process. Here, we describe a yeast-recombination-based approach to construct and mutate plasmids containing the cDNA of the human bile salt export pump (BSEP) that has been shown to be unstable in E. coli. Using this approach, we constructed the necessary plasmids for a heterologous overexpression of BSEP in the yeast Pichia pastoris. We then applied a new site-directed mutagenesis method, DREAM (Directed REcombination-Assisted Mutagenesis) that completely bypasses E. coli by using S. cerevisiae as the plasmid host with high mutagenesis efficiency. Finally, we show how to apply this strategy to unstable non-yeast plasmids by rapidly turning an existing mammalian BSEP expression construct into a S. cerevisiae-compatible plasmid and analyzing the impact of a BSEP mutation in several mammalian cell lines