Subcellular imaging of intramitochondrial Ca2+ with recombinant targeted aequorin: significance for the regulation of pyruvate dehydrogenase activity.

Specific targeting of the recombinant, Ca2+ -sensitive photoprotein, aequorin to intracellular organelles has provided new insights into the mechanisms of intracellular Ca2+ homeostasis. When applied to small mammalian cells, a major limitation of this technique has been the need to average the signal over a large number of cells. This prevents the identification of inter- or intracellular heterogeneities. Here we describe the imaging in single mammalian cells (CHO.T) of [Ca2+] with recombinant chimeric aequorin targeted to mitochondria. This was achieved by optimizing expression of the protein through intranuclear injection of cDNA and through the use of a charge-coupled device camera fitted with a dual microchannel plate intensifier. This approach allows accurate quantitation of the kinetics and extent of the large changes in mitochondrial matrix [Ca2+] ([Ca2+](m)) that follow receptor stimulation and reveal different behaviors of mitochondrial populations within individual cells. The technique is compared with measurements of [Ca2+](m) using the fluorescent indicator, rhod2. Comparison of [Ca2+](m) with the activity of the Ca2+ -sensitive matrix enzyme, pyruvate dehydrogenase (PDH), reveals that this enzyme is a target of the matrix [Ca2+] changes. Peak [Ca2+](m) values following receptor stimulation are in excess of those necessary for full activation of PDH in situ, but may be necessary for the activation of other mitochondrial dehydrogenases. Finally, the data suggest that the complex regulation of PDH activity by a phosphorylation-dephosphorylation cycle may provide a means by which changes in the frequency of cytosolic (and hence mitochondrial) [Ca2+] oscillations can be decoded by mitochondria.

Significance of bound water to local chain conformations in protein crystals.

We examine how the polypeptide chain in protein crystal structures exploits the multivalent hydrogen-bonding potential of bound water molecules. This shows that multiple interactions with a single water molecule tend to occur locally along the chain. A distinctive internal-coordinate representation of the local water-binding segments reveals several consensus conformations. The fractional water occupancy of each was found by comparison of the total number of conformations in the database regardless of the presence or absence of bound water. The water molecule appears particularly frequently in type II beta-turn geometries and an N-terminal helix feature. This work constitutes a first step into assessing not only the generality but also the significance of specific water binding in globular proteins.

Brachiopod assemblages from the Early-Middle Jurassic transition in the Eastern Subbetic (SE Spain): Systematic and palaeobiogeographic implications and palaeoenvironmental significance

New data on brachiopod assemblages recorded in the Eastern Subbetic area (Alicante, SE Spain) and attributed to the Early Bajocian (Humphriesianum Zone and/or immediately older) are provided. Eleven species have been distinguished and reported for the first time in the Subbetic domain of the Betic Cordillera. The description of the morphological evidences on each analysed taxa, especially in relation to their internal morphology, brings new implications in the systematics of the Middle Jurassic brachiopods. The analysis of faunistic affinity between the recorded assemblages and those from other palaeogeographic domains, shows that the Subbetic brachiopod fauna has a clear Mediterranean affinity, as proved by the different species belonging to the genera Striirhynchia, Septocrurella, Mondegia?, Karadagithyris, Linguithyris, Papodina?, Viallithyris?, and Zugmayeria?. It is also evidenced that the Early-Middle Jurassic transition in the Eastern Subbetic accounted, in qualitative terms, a remarkable interval of faunistic renewal in the brachiopod assemblages, strongly influenced by a complex tectonic and stratigraphic framework controlled by a period of intense extensional tectonics, globally framed in the evolution of the Atlantic Ocean.

The functional significance of the hypocercal tail and lateral fin fold of Anaspid ostracoderms / James A. Hopson --.

v.33:no.5(1974)

Fossil shrews from Honduras and their significance for late glacial evolution in body size (Mammalia: Soricidae: Cryptotis) / Neal Woodman --, Darin A. Croft --.

n.s. no.51(2005)

Ethnological and biogeographical significance of pottery sherds from Nissan Island, Papua New Guinea / Susan Kaplan, Bryn Mawr College.

v.66:no.3(1976)

Some Middle Pennsylvanian Hoplocarida (Crustacea) and their phylogenetic significance / Frederick R. Schram --

v.12:no.14(1969)

The Significance of the Disputed Territory in Bechuanaland 1887-1893

British Imperial policy in Southern Africa in the last three decades of the nineteenth century oscillated between two extremes. It began in the early 1870's with Lord Kimberley's attempt to effect confederation as a means of devolving Imperial responsibility and expenditure. It ended in 1899 with Britain's active intervention against the Boers. For most of the remaining years of those decades a middle course was adopted while the British Government struggled to reconcile its diverse political interests. Strategy, supremacy, economy, humanitarianism, and recognition of colonial aspirations were all at one time or another, in varying degrees, motivating forces behind Imperial policy. Many historians have pointed out how incompatible many of these ends were and how the attempt to pursue them all at once almost inevitably ended in at least one of them being sacrificed on the way. This study focusses on a relatively minor problem over a period of about seven years. It attempts to show how the British Government tried to reconcile, in this case, the predominant motives of economy and supremacy. The problem of the Disputed Territory now seems like a small fish in a big ocean because non the great hopes and fears that it raised were ever realized. But the anticlimactic nature of the outcome of events should not be allowed to conceal two important points: first, that the problem loomed large at the time in the eyes of the Imperial Government; and second, that in the case of its policy towards the Disputed Territory, the Government gained a greater degree of success in trying to reconcile seemingly incompatible ends than it did in many other instances.

The significance of EU topics in national media. Has there been an Europeanization of reporting in the national media? Bruges Political Research Papers No. 27/November 2012

Criticisms are often voiced at the fact that there is no well-informed European public. However, as the process of European integration has advanced, the media have been devoting more resources and space to the coverage of European affairs. At the same time, the national media have gone from being mere transmitters of information to having their own voice on European issues. In this respect, the media have emerged as actors capable of influencing the opinions of citizens, thereby contributing to the emergence of a European public sphere. The present study analyzes whether a Europeanization of the national media has taken place by studying how national newspapers provide information in Europe and whether a European public sphere is emerging. The results reveal that some European topics have experienced a certain Europeanization, but there is still an absence of European debate in the respective national public spheres.

The "European model" of international law and its significance for Asia. Some critical reflections. EU Centre in Singapore Working Paper No. 7, June 2012

There is a certain arrogance in the affirmation that a “European model” of regional integration and of compliance with international law should be adopted anywhere in the world, and in Asia in particular. This article argues on the contrary that Asia and Europe are in fundamentally different situations vis-à-vis international law. Based on an analysis of recent events and latest legal developments in Europe, it puts the “European model” of regional integration and the European selective compliance with international law in perspective with regard to the Asian context. Without denying that “civilizations” should learn from one another and that the European experience may be relevant to some extent in Asia, this article concludes that the tools developed in Europe should be used differently in Asia.