Neuronal precursor cell-expressed developmentally down-regulated 4-1 (NEDD4-1) controls the sorting of newly synthesized Ca(V)1.2 calcium channels

Neuronal precursor cell-expressed developmentally down-regulated 4 (Nedd4) proteins are ubiquitin ligases, which attach ubiquitin moieties to their target proteins, a post-translational modification that is most commonly associated with protein degradation. Nedd4 ubiquitin ligases have been shown to down-regulate both potassium and sodium channels. In this study, we investigated whether Nedd4 ubiquitin ligases also regulate Ca(v) calcium channels. We expressed three Nedd4 family members, Nedd4-1, Nedd4-2, and WWP2, together with Ca(v)1.2 channels in tsA-201 cells. We found that Nedd4-1 dramatically decreased Ca(v) whole-cell currents, whereas Nedd4-2 and WWP2 failed to regulate the current. Surface biotinylation assays revealed that Nedd4-1 decreased the number of channels inserted at the plasma membrane. Western blots also showed a concomitant decrease in the total expression of the channels. Surprisingly, however, neither the Ca(v) pore-forming α1 subunit nor the associated Ca(v)β and Ca(v)α(2)δ subunits were ubiquitylated by Nedd4-1. The proteasome inhibitor MG132 prevented the degradation of Ca(v) channels, whereas monodansylcadaverine and chloroquine partially antagonized the Nedd4-1-induced regulation of Ca(v) currents. Remarkably, the effect of Nedd4-1 was fully prevented by brefeldin A. These data suggest that Nedd4-1 promotes the sorting of newly synthesized Ca(v) channels for degradation by both the proteasome and the lysosome. Most importantly, Nedd4-1-induced regulation required the co-expression of Ca(v)β subunits, known to antagonize the retention of the channels in the endoplasmic reticulum. Altogether, our results suggest that Nedd4-1 interferes with the chaperon role of Ca(v)β at the endoplasmic reticulum/Golgi level to prevent the delivery of Ca(v) channels at the plasma membrane.

Climate warming and vegetation response after Heinrich event 1 (16 700–16 000 cal yr BP) in Europe south of the Alps

Chironomids preserved in a sediment core from Lago di Origlio (416 m a.s.l.), a lake in the foreland of the Southern Swiss Alps, allowed quantitative reconstruction of Late Glacial and Early Holocene summer temperatures using a combined Swiss–Norwegian temperature inference model based on chironomid assemblages from 274 lakes. We reconstruct July air temperatures of ca. 10 °C between 17 300 and 16 000 cal yr BP, a rather abrupt warming to ca. 12.0 °C at ca. 16 500–16 000 cal yr BP, and a strong temperature increase at the transition to the Bølling/Allerød interstadial with average temperatures of about 14 °C. During the Younger Dryas and earliest Holocene similar temperatures are reconstructed as for the interstadial. The rather abrupt warming at 16 500–16 000 cal yr BP is consistent with sea-surface temperature as well as speleothem records, which indicate a warming after the end of Heinrich event 1 (sensu stricto) and before the Bølling/Allerød interstadial in southern Europe and the Mediterranean Sea. Pollen records from Origlio and other sites in southern Switzerland and northern Italy indicate an early reforestation of the lowlands 2000–1500 yr prior to the large-scale afforestation of Central Europe at the onset of the Bølling/Allerød period at ca. 14 700–14 600 cal yr BP. Our results suggest that these early afforestation processes in the formerly glaciated areas of northern Italy and southern Switzerland have been promoted by increasing temperatures.

Mechanism of Ca(v)1.2 channel modulation by the amino terminus of cardiac beta2-subunits

L-type calcium channels are composed of a pore, alpha1c (Ca(V)1.2), and accessory beta- and alpha2delta-subunits. The beta-subunit core structure was recently resolved at high resolution, providing important information on many functional aspects of channel modulation. In this study we reveal differential novel effects of five beta2-subunits isoforms expressed in human heart (beta(2a-e)) on the single L-type calcium channel current. These splice variants differ only by amino-terminal length and amino acid composition. Single-channel modulation by beta2-subunit isoforms was investigated in HEK293 cells expressing the recombinant L-type ion conducting pore. All beta2-subunits increased open probability, availability, and peak current with a highly consistent rank order (beta2a approximately = beta2b > beta2e approximately = beta2c > beta2d). We show graded modulation of some transition rates within and between deep-closed and inactivated states. The extent of modulation correlates strongly with the length of amino-terminal domains. Two mutant beta2-subunits that imitate the natural span related to length confirm this conclusion. The data show that the length of amino termini is a relevant physiological mechanism for channel closure and inactivation, and that natural alternative splicing exploits this principle for modulation of the gating properties of calcium channels.

Ubiquitin-specific protease USP2-45 acts as a molecular switch to promote α2δ-1-induced downregulation of Ca v1.2 channels

Availability of voltage-gated calcium channels (Cav) at the plasma membrane is paramount to maintaining the calcium homeostasis of the cell. It is proposed that the ubiquitylation/de-ubiquitylation balance regulates the density of ion channels at the cell surface. Voltage-gated calcium channels Cav1.2 have been found to be ubiquitylated under basal conditions both in vitro and in vivo. In a previous study, we have shown that Cav1.2 channels are ubiquitylated by neuronal precursor cell-expressed developmentally downregulated 4 (Nedd4-1) ubiquitin ligases, but the identity of the counterpart de-ubiquitylating enzyme remained to be elucidated. Regarding sodium and potassium channels, it has been reported that the action of the related isoform Nedd4-2 is counteracted by the ubiquitin-specific protease (USP) 2-45. In this study, we show that USP 2-45 also de-ubiquitylates Cav channels. We co-expressed USPs and Cav1.2 channels together with the accessory subunits β2 and α2δ-1, in tsA-201 and HEK-293 mammalian cell lines. Using whole-cell current recordings and surface biotinylation assays, we show that USP2-45 specifically decreases both the amplitude of Cav currents and the amount of Cav1.2 subunits inserted at the plasma membrane. Importantly, co-expression of the α2δ-1 accessory subunit is necessary to support the effect of USP2-45. We further show that USP2-45 promotes the de-ubiquitylation of both Cav1.2 and α2δ-1 subunits. Remarkably, α2δ-1, but not Cav1.2 nor β2, co-precipitated with USP2-45. These results suggest that USP2-45 binding to α2δ-1 promotes the de-ubiquitylation of both Cav1.2 and α2δ-1 subunits, in order to regulate the expression of Cav1.2 channels at the plasma membrane.

Tab. 1+2: C14-dates cf the older and younger emergence periods

In the neighbourhood of Oobloyah Bay various phenomena ean be eneountered whieh point to a ularge-seale uplift of shorelines, i.e. to an emergence of 200 m. Delta terraces, deltaic fan terraces and glacio-marine sands are regarded by the author as being the most reliable evidence of this. The marine limit documented by glacio-marine sand is to be found at ~170 m a.s.l. Hints of ancient shorelines located at a higher level exist only in the shape of badly preserved raised beaches. They were classified as less reliable records of past sea-levels, due to the lack of marine fossils and/or drift wood, and furthermore because those forms had been strongly influenced by periglacial processes. Deltaic deposits are of more importance in this context. The glacio-marine deltaic sands of several terrace levels contain terrestrial plant remnants which delivered C14dates. Using these dates und the relative elevations of terraces the emergenee of the area investigated could be recorded. This occured in a series of phases (and steps) which were summarized into two periods: an early period of emergenee which took place from at least 25 300 years B.P. to later than 17 340 years B.P. and a later one from at least 12 870 years B.P. up to the present day. The emergence seems to represent a discontinuous but regular sequence of relative sea level movements without intermittent submergence. Since the deltaic fans of the early emergence period were accumulated by sediments through glacio-fluvial channels of an adjacent glacier body the appropriate location of this glacial stage for one of the glaciers delivering meltwater (Nukapingwa Glacier) could be reconstructed. This stage of the glacier appears to belong to a retreating phase of the Mid-Wisconsin (?). The later period of emergence resulted in six rather glacio-marine delta terrace generations at the mouths of the main rivers with glaciofluvial regimen debouching into the Oobloyah Bay. A connection of this emergence with the glacial history of the field area is discussed. If one may rely on the age determinations of land derived plant fossils and their application for the climatic history of the area investigated, it must be concluded that the Heidelberg Valley, to a large extent, was alreaely deglaciated 25 000 years ago. The existence of a "Cockburn"-Phase in the sense of a major readvanee in Late Wisconsin times appears to be doubtful, or has been developed rather weakly.

A foraminiferal d18O record of sediment core GT90-3 covering the last 2,200 years

For many years the Torino Cosmogeophysics group has been studying sediment cores drilled from the Gallipoli Terrace in the Gulf of Taranto (Ionian Sea) and deposited in the last millennia. The gravity core GT90-3, in which the 18O series was measured, was drilled from the Gallipoli Terrace in the Gulf of Taranto (Ionian Sea) at 39°45'53''N, 17°53'33''E. It was extracted at a depth of 178 m and its length is 3.57 m. Thanks to its geographical location, the Gallipoli Terrace is a favourable site for climatic studies based on marine sediments, because of its closeness to the volcanically active Campanian area, a region that is unique in the world for its detailed historical documentation of volcanic eruptions. Tephra layers corresponding to historical eruptions were identified along the cores, thus allowing for accurate dating and determination of the sedimentation rate. The measurements performed in different cores from the same area showed that the sedimentation rate is uniform across the whole Gallipoli Terrace. We measured the oxygen isotope composition d18O of planktonic foraminifera. These measurements provided a high-resolution 2,200-year-long record. We sampled the core using a spacing of 2.5 mm corresponding to 3.87 years. Each sample of sediment (5 g) was soaked in 5% calgon solution overnight, then treated in 10% H2O2 to remove any residual organic material. Subsequently it was washed with a distilled-water jet through a sieve with a 150 µm mesh. The fraction > 150 µm was kept and oven-dried at 5°C. The planktonic foraminifera Globigerinoides ruber were picked out of the samples under a microscope. For each sample, 20-30 specimens were selected from the fraction comprised between 150 µm and 300 µm. The use of a relatively large number of specimens for each sample reduces the isotopic variability of individual organisms, giving a more representative d18O value. The stable isotope measurements were performed using a VG-PRISM mass spectrometer fitted with an automated ISO-CARB preparation device. Analytical precision based on internal standards was better than 0.1 per mil. Calibration of the mass spectrometer to VPDB scale was done using NBS19 and NBS18 carbonate standards. The strategic location of the drilling area makes this record a unique tool for climate and oceanographic studies of the Central Mediterranean.

Propagation of Cross-Section Uncertainties in Criticality Calculations in the Framework of UAM-Phase I Using MCNPX-2.7e and SCALE-6.1

In the framework of the OECD/NEA project on Benchmark for Uncertainty Analysis in Modeling (UAM) for Design, Operation, and Safety Analysis of LWRs, several approaches and codes are being used to deal with the exercises proposed in Phase I, “Specifications and Support Data for Neutronics Cases.” At UPM, our research group treats these exercises with sensitivity calculations and the “sandwich formula” to propagate cross-section uncertainties. Two different codes are employed to calculate the sensitivity coefficients of to cross sections in criticality calculations: MCNPX-2.7e and SCALE-6.1. The former uses the Differential Operator Technique and the latter uses the Adjoint-Weighted Technique. In this paper, the main results for exercise I-2 “Lattice Physics” are presented for the criticality calculations of PWR. These criticality calculations are done for a TMI fuel assembly at four different states: HZP-Unrodded, HZP-Rodded, HFP-Unrodded, and HFP-Rodded. The results of the two different codes above are presented and compared. The comparison proves a good agreement between SCALE-6.1 and MCNPX-2.7e in uncertainty that comes from the sensitivity coefficients calculated by both codes. Differences are found when the sensitivity profiles are analysed, but they do not lead to differences in the uncertainty.

Transvaal & Orange Free State Regions, South Africa, ca. 1899 (Raster Image)

This layer is a georeferenced raster image of the historic paper map entitled: Bacon's large-print map of the Transvaal and Orange Free State. It was published by G.W. Bacon & Co. ca. 1899. Scale [ca. 1:1,900,000]. Covers also Swaziland, Lesotho, and portions of Botswana, Zimbabwe, and Mozambique.The image inside the map neatline is georeferenced to the surface of the earth and fit to the Africa Sinusoidal projected coordinate system. All map collar and inset information is also available as part of the raster image, including any inset maps, profiles, statistical tables, directories, text, illustrations, index maps, legends, or other information associated with the principal map. This map shows features such as drainage, cities and other human settlements, territorial and administrative boundaries, roads, railroads, shoreline features, and more. Relief shown by shading and spot heights. Includes also insets: "Map showing the routes from England and India to South Africa", "Environs of Cape Town", "Lorenço Marquez [and vicinity]", 'South Africa" and "Durban and Port Natal".This layer is part of a selection of digitally scanned and georeferenced historic maps from the Harvard Map Collection. These maps typically portray both natural and manmade features. The selection represents a range of originators, ground condition dates, scales, and map purposes.

Jamaica, 1763 (Image 3 of 4) (Raster Image)

This layer is a georeferenced raster image of the historic paper map entitled: This map of the island of Jamaica; laid down from the papers and under the direction of Henry Moore, Esqr.; His Majesty's Lieutenant Governor and Commander in Chief of that island, in the years 1756, 57, 58, 59, 60 & 61; & from a great number of actual surveys performed by the publishers is humbly inscribed by his lordship's most obedient & most humble servants, Thos. Craskell, engineer, Jas. Simpson, surveyor. It was published by D. Fournier in 1763. Scale [ca 1:200,000]. This layer is image 3 of 4 total images of the four sheet map, representing the northeast portion of the map. The image inside the map neatline is georeferenced to the surface of the earth and fit to the Jamaica Grid projected coordinate system. All map collar and inset information is also available as part of the raster image, including any inset maps, profiles, statistical tables, directories, text, illustrations, index maps, legends, or other information associated with the principal map. This map shows features such as towns, villages, and other human settlements, roads, drainage, selected buildings, ground cover, shoreline features, and more. Relief shown pictorially. Includes also illustrations.This layer is part of a selection of digitally scanned and georeferenced historic maps from the Harvard Map Collection. These maps typically portray both natural and manmade features. The selection represents a range of originators, ground condition dates, scales, and map purposes.

Jamaica, 1763 (Image 4 of 4) (Raster Image)

This layer is a georeferenced raster image of the historic paper map entitled: This map of the island of Jamaica; laid down from the papers and under the direction of Henry Moore, Esqr.; His Majesty's Lieutenant Governor and Commander in Chief of that island, in the years 1756, 57, 58, 59, 60 & 61; & from a great number of actual surveys performed by the publishers is humbly inscribed by his lordship's most obedient & most humble servants, Thos. Craskell, engineer, Jas. Simpson, surveyor. It was published by D. Fournier in 1763. Scale [ca 1:200,000]. This layer is image 4 of 4 total images of the four sheet map, representing the northwest portion of the map. The image inside the map neatline is georeferenced to the surface of the earth and fit to the Jamaica Grid projected coordinate system. All map collar and inset information is also available as part of the raster image, including any inset maps, profiles, statistical tables, directories, text, illustrations, index maps, legends, or other information associated with the principal map. This map shows features such as towns, villages, and other human settlements, roads, drainage, selected buildings, ground cover, shoreline features, and more. Relief shown pictorially. Includes also illustrations.This layer is part of a selection of digitally scanned and georeferenced historic maps from the Harvard Map Collection. These maps typically portray both natural and manmade features. The selection represents a range of originators, ground condition dates, scales, and map purposes.

County of Cornwall, Jamaica, 1763 (Image 1 of 4) (Raster Image)

This layer is a georeferenced raster image of the historic paper map entitled: This map of the county of Cornwall, in the island of Jamaica; laid down from the papers, under the direction of, Henry Moore, Esqr., His Majesty's Lieutenant Governor, and Commander in Chief of that island, in the years 1756, 57, 58, 59, 60 & 61, & from a great number of actual surveys performed by the publishers is humbly inscribed by his lordship's most obedient & humble servants, Thos. Craskell, engineer, Jas. Simpson, surveyor. It was published by D. Fournier in 1763. Scale [ca. 1:95,000]. This layer is image 1 of 4 total images of the four sheet source map, representing the southwest portion of the map. The image inside the map neatline is georeferenced to the surface of the earth and fit to the Jamaica Grid projected coordinate system. All map collar and inset information is also available as part of the raster image, including any inset maps, profiles, statistical tables, directories, text, illustrations, index maps, legends, or other information associated with the principal map. This map shows features such as towns, villages, and other human settlements, roads, parish boundaries, drainage, selected buildings and names of landowners, ground cover, shoreline features, shoals, channels, anchorage points, and more.This layer is part of a selection of digitally scanned and georeferenced historic maps from the Harvard Map Collection. These maps typically portray both natural and manmade features. The selection represents a range of originators, ground condition dates, scales, and map purposes.