Alterations in the cytoplasmic domain of CLCN2 result in altered gating kinetics.

Mutations in the human ClC-2 Cl(-) channel have been described to influence its function dramatically. To test for naturally occurring gene variants in a human population and their functionality, all 24 CLCN2 exons from a Central African population were sequenced. Six single amino acid exchanges in the intracellular N-terminus (P48R, R68H), in the pore domain (G199A), or in the intracellular C-terminus (R646Q, R725W, R747H) were identified at low frequency. Heterologous expression of these polymorphisms in Xenopus laevis oocytes demonstrated their functional significance as determined by two-electrode voltage-clamp. The polymorphisms R68H, R725W, and R747H exhibited faster voltage-stimulated gating as compared to the wild type channel, resulting in higher steady state currents of R725W. Probably due to decreased surface expression P48R, R68H, and R646Q mutants generated lower currents than the wild type channels. The inward currents of the mutated channels R725W, R747H, and G199A failed to increase during hypotonic swelling, a defect paralleled by impaired swelling-accelerated voltage-gating in one mutant (G199A). In conclusion, the Africans' gene pool comprises CLCN2 gene variants in the N-terminus, the C-terminus or the pore domain that affect surface expression and voltage- or cell-swelling-stimulated channel gating.     

Avian Hepatitis B Virus Infection Is Initiated by the Interaction of a Distinct Pre-S Subdomain with the Cellular Receptor gp180

Functionally relevant hepadnavirus-cell surface interactions were investigated with the duck hepatitis B virus (DHBV) animal model by using an in vitro infection competition assay. Recombinant DHBV pre-S polypeptides, produced in Escherichia coli, were shown to inhibit DHBV infection in a dose-dependent manner, indicating that monomeric pre-S chains were capable of interfering with virus-receptor interaction. Particle-associated pre-S was, however, 30-fold more active, suggesting that cooperative interactions enhance particle binding. An 85-amino-acid pre-S sequence, spanning about half of the DHBV pre-S chain, was characterized by deletion analysis as essential for maximal inhibition. Pre-S polypeptides from heron hepatitis B virus (HHBV) competed DHBV infection equally well despite a 50% difference in amino acid sequence and a much-reduced infectivity of HHBV for duck hepatocytes. These observations are taken to indicate (i) that the functionality of the DHBV pre-S subdomain, which interacts with the cellular receptor, is determined predominantly by a defined three-dimensional structure rather than by primary sequence elements; (ii) that cellular uptake of hepadnaviruses is a multistep process involving more than a single cellular receptor component; and (iii) that gp180, a cellular receptor candidate unable to discriminate between DHBV and HHBV, is a common component of the cellular receptor complex for avian hepadnaviruses.     

Tectonic and volcanic controls on Early Jurassic rift-valley lake deposition during emplacement of Karoo flood basalts, southern Namibia

The Karoo Igneous Province of southern Africa is one of the classic Mesozoic flood basalt provinces of the world. In the case of the early Jurassic Kalkrand Formation of Namibia the succession comprises three major flood basalt units that are separated by two stratigraphically important fluvio-lacustrine interlayers. These horizons preserve a record of the complex interplay between sedimentation, effusion of Karoo flood basalts and extensional tectonics that predated and accompanied the break-up of Gondwanaland. Both sediment layers start with the dominantly local derivation of weathered and eroded lava debris, followed by the emplacement of subaqueous mass flows and subsequent deposition of chemical sediments. The latter are characterised by interbedded stromatolitic carbonates, grass-like structured gypsum, and plane-bedded sandstones and mudstones containing euhedral displacive gypsum crystals that grew in the subsurface as well as rosettes which nucleated on the sediment surface. The central parts of the lacustrine units are overlain by thin deltaic sandstones showing bottomset, foreset, and topset layering and, finally, braided fluvial, trough cross-bedded sandstones. Evidence of subsidence synchronous with the formation of lake bodies can be explained by two principal mechanisms. The first acted in localised areas only and is reflected by the development of small, centrally subsiding basins. From the repeated occurrence of onlapping geometries within such a pool, a multiphase history of sagging is deduced, being most likely related to periodic magma withdrawal and reduction in magmatic pressures in subsurface lava feeders beneath the basin floor. Abundant syn-subsident fracture features at lava–sediment contacts, such as sediment-, hydrothermal calcite-, and sometimes basic lava-filled fissure systems indicate the pronounced interaction between the underlying volcanics and these small areas of pronounced subsidence. Fluids passing through the volcanic pile exhaled into the lake, giving it the characteristics of alkaline lake systems described from more recent flood basalt areas associated with the modern African Rift System. On the regional scale, however, northerly trending extensional fault systems controlled half-graben basin geometries and both facies and thickness variations across faults indicate that tectonism operated contemporaneously with volcanism and lacustrine sedimentation. The analysis of faults and associated structures, such as regularly aligned sediment-filled fissures, sets of micro-faults, folds and basaltic dykes constrains the extensional opening direction for the Karoo graben structures in this area that heralded the opening of the South Atlantic and thus provides a basis to discuss the extensional history of the Namibian coastal margin within the regional tectonic framework.     

Cell-Dependent Differences in the Production of Infectious Herpes Simplex Virus at a Supraoptimal Temperature

The replication of herpes simplex virus (HSV) was compared in rabbit and hamster cells at optimal and supraoptimal temperatures. Replication occurred in cells of either species at 33 C, but the total infectious virus yield was routinely about 10-fold greater in rabbit cells than in hamster cells. At 39 C, this difference was exaggerated to greater than 100,000-fold. Whereas infectious virus was produced and plaques formed in rabbit kidney cell monolayers at the higher temperature, neither developed in those derived from hamster embryos. Elevating the temperature from 33 C to 39 C at various time intervals after exposure of the cultures to virus revealed that production of infectious virus in hamster cells was completely heat-sensitive up to 6 hr after infection. Specific viral antigens and viral deoxyribonucleic acid (DNA) were synthesized in both rabbit and hamster cell cultures. In addition, cellular DNA synthesis was depressed and cytopathic effects occurred in both cell systems. These cytopathic effects were not observed in cell cultures treated with HSV previously inactivated with ultraviolet light. Compared with parallel cultures at 33 C, the amount of viral DNA synthesized at 39 C was greatly reduced in both systems. In hamster cells, the reduction was twofold greater than in rabbit cells. This cell-dependent thermal inhibition of HSV replication in hamster cells did not occur with vaccinia virus.     

Activation of a Latent Measles Virus Infection in Hamster Cells

The characteristics of infectious measles virus released from latently infected hamster embryo fibroblast cells are described. Low levels of virus were released spontaneously when the cultures were incubated at 37 C; this phenomenon was observed 19 passages after the cells had been exposed to the virus and has continued through cell passage 45. The virus yield could be significantly increased by cocultivation of the hamster cells with BSC-1 cells or incubation of the latently infected cells at 33.5 C rather than at 37 C. Measles virus released after cocultivation demonstrated increased cytopathology in cell culture and reduced temperature sensitivity when compared to the virus released at 33.5 C. After cell passage 45, there was an increase in spontaneous release of virus. However, the viruses recovered by cocultivation or temperature release after cell passage 45 were nearly identical. These observations suggest a possible mechanism for measles virus activation in cells latently infected with this virus.     

Mykologisches

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