Perforin is a secreted protein synthesized by activated cytotoxic T lymphocytes (CTL) and natural killer (NK) cells. It is a key component of the lytic machinery of these cells, being able to insert into the plasma membrane of targeted cells, forming a pore which leads to their destruction. Here we analyse the synthesis, processing and intracellular transport of perforin in the NK cell line YT. Perforin is synthesized as a 70 kDa inactive precursor which is cleaved at the C-terminus to yield a 60 kDa active form. This proteolytic cleavage occurs in an acidic compartment and can be inhibited by incubation of the cells in ammonium chloride, concanamycin A, leupeptin and E-64. The increased lytic activity of the cleaved form can be demonstrated by killing assays in which cleavage of the pro-piece is inhibited. Epitope mapping reveals that cleavage of the pro-piece occurs at the boundary of a C2 domain, which we show is able to bind phospholipid membranes in a calcium-dependent manner. We propose that removal of the pro-piece, which contains a bulky glycan, allows the C2 domain to interact with phospholipid membranes and initiate perforin pore formation.

Suitable thermal conditions in streams are necessary for fish and predictions of future climate changes infer that water temperatures may regularly exceed tolerable ranges for key species. Riparian woodland is considered as a possible management tool for moderating future thermal conditions in streams for the benefit of fish communities. The spatial and temporal variation of stream water temperature was therefore investigated over 3 years in lowland rivers in the New Forest (southern England) to establish the suitability of the thermal regime for fish in relation to riparian shade in a warm water system. Riparian shade was found to have a marked influence on stream water temperature, particularly in terms of moderating diel temperature variation and limiting the number of days per year that maximum temperatures exceeded published thermal thresholds for brown trout. Expansion of riparian woodland offers potential to prevent water temperature exceeding incipient lethal limits for brown trout and other fish species. A relatively low level of shade (20-40%) was found to be effective in keeping summer temperatures below the incipient lethal limit for brown trout, but ca. 80% shade generally prevented water temperatures exceeding the range reported for optimum growth of brown trout. Higher levels of shade are likely to be necessary to protect temperature-sensitive species from climate warming. # Crown copyright 2010.

The appearance of farming, from its inception in the Near East around 12 000 years ago, finally reached the northwestern extremes of Europe by the fourth millennium BC or shortly thereafter. Various models have been invoked to explain the Neolithization of northern Europe; however, resolving these different scenarios has proved problematic due to poor faunal preservation and the lack of specificity achievable for commonly applied proxies. Here, we present new multi-proxy evidence, which qualitatively and quantitatively maps subsistence change in the northeast Atlantic archipelagos from the Late Mesolithic into the Neolithic and beyond. A model involving significant retention of hunter–gatherer–fisher influences was tested against one of the dominant adoptions of farming using a novel suite of lipid biomarkers, including dihydroxy fatty acids, ω-(o-alkylphenyl)alkanoic acids and stable carbon isotope signatures of individual fatty acids preserved in cooking vessels. These new findings, together with archaeozoological and human skeletal collagen bulk stable carbon isotope proxies, unequivocally confirm rejection of marine resources by early farmers coinciding with the adoption of intensive dairy farming. This pattern of Neolithization contrasts markedly to that occurring contemporaneously in the Baltic, suggesting that geographically distinct ecological and cultural influences dictated the evolution of subsistence practices at this critical phase of European prehistory.

Methodological advances in dating the Middle to Upper Paleolithic transition provide a better understanding of the replacement of local Neanderthal populations by Anatomically Modern Humans. Today we know that this replacement was not a single, pan-European event, but rather it took place at different times in different regions. Thus, local conditions could have played a role. Iberia represents a significant macro-region to study this process. Northern Atlantic Spain contains evidence of both Mousterian and Early Upper Paleolithic occupations, although most of them are not properly dated, thus hindering the chances of an adequate interpretation. Here we present 46 new radiocarbon dates conducted using ultrafiltration pre-treatment method of anthropogenically manipulated bones from 13 sites in the Cantabrian region containing Mousterian, Aurignacian and Gravettian levels, of which 30 are considered relevant. These dates, alongside previously reported ones, were integrated into a Bayesian age model to reconstruct an absolute timescale for the transitional period. According to it, the Mousterian disappeared in the region by 47.9–45.1ka cal BP, while the Châtelperronian lasted between 42.6k and 41.5ka cal BP. The Mousterian and Châtelperronian did not overlap, indicating that the latter might be either intrusive or an offshoot of the Mousterian. The new chronology also suggests that the Aurignacian appears between 43.3–40.5ka cal BP overlapping with the Châtelperronian, and ended around 34.6–33.1ka cal BP, after the Gravettian had already been established in the region. This evidence indicates that Neanderthals and AMH co-existed <1,000 years, with the caveat that no diagnostic human remains have been found with the latest Mousterian, Châtelperronian or earliest Aurignacian in Cantabrian Spain.

The rate of release of Fe(I1) from anoxic lake sediments was lower in the presence than in the absence of nitrate. The reduction of Fe(II1) by the sediments had a temperature optimum of 30 "C and was inhibited by HgC12, suggesting that the process was largely biological in nature.Of the iron sources tested with cultures of anaerobic iron-reducing bacteria, FeC13 was the most readily reduced and goethite the least. Reduction was faster in the presence of a chelating agent and was suppressed by the addition of NO,, C105, MnO,, Mn203 and 02. An iron-reducing chemoorganotroph, tentatively identified as a member of the genus Vibrio, was isolated. Physical contact between the bacterium and iron particles was essential to ensure maximum rates of Fe(II1) reduction but > 30% of the activity appeared to be associated with extracellular components. Although Fe(II1) reduction by whole cells and cell-free extracts was decreased in the presence of electron transport inhibitors, the molar growth yield of the organism was unaffected by the presence of Fe(II1). It is assumed that the organism used the Fe(II1) as a hydrogen sink. A second organism, an anaerobic facultative chemolithotroph, appeared to conserve energy by the reduction of Fe(II1). Biomass yield (measured as ATP) was greater in the presence of Fe(III), and the organism was able to use H2 as a source of reducing power.