Isolation, characterization and functional analysis of the {\it xnf7\/} gene from {\it Xenopus laevis\/}

A fundamental problem in developmental biology concerns the mechanisms involved in the establishment of the embryonic axis. We are studying Xenopus nuclear factor 7 (xnf7) which we believe to be involved in dorsal-ventral patterning in Xenopus laevis. Xnf7 is a maternal gene product that is retained in the cytoplasm during early embryogenesis until the mid-blastula transition (MBT) when it reenters the nuclei. It is a member of a novel zinc finger proteins, the B-box family, consisting mainly of transcription factors and protooncogenes.^ The xnf7 gene is reexpressed during embryogenesis at the gastrula-neurula stage of development, with its zygotic expression limited to the central nervous system (CNS). In this study we showed that there are two different cDNAs coding for xnf7, xnf7-O and xnf7-B. They differ by 39 amino acid changes scattered throughout the cDNA. The expression of both forms of xnf7 is limited primarily to the central nervous system (CNS) and dorsal axial structures during later stages of embryogenesis.^ In order to study the spatial and temporal regulation of the gene, we screened a Xenopus genomic library using part of xnf7 cDNA as a probe. A genomic clone corresponding to the xnf7-O type was isolated, its 5$\sp\prime$ putative regulatory region sequenced, and its transcriptional initiation site mapped. The putative promoter region contained binding sites for Sp1, E2F, USF, a Pu box and AP1. CAT/xnf7 fusion genes were constructed containing various 5$\sp\prime$ deleted regions of the xnf7 promoter linked to a CAT (Chloramphenicol Acetyl Transferase) reporter vector. These constructs were injected into Xenopus oocytes and embryos to study the regions of the xnf7 promoter responsible for basal, temporal and spatial regulation of the gene. The activity of the fusion genes was measured by the conversion of chloramphenicol to its acetylated forms, and the spatial distribution of the transcripts by whole mount in situ hybridization. We showed that the elements involved in basal regulation of xnf7 lie within 121 basepairs upstream of the transcriptional inititiation site. A DNase I footprint analysis performed using oocyte extract showed that a E2F and 2 Sp1 sites were protected. During development, the fusion genes were expressed following the MBT, in accordance with the timing of the endogenous xnf7 gene. Spatially, the expression of the fusion gene containing 421 basepairs of the promoter was localized to the dorsal region of the embryo in a pattern that was almost identical to that detected with the endogenous transcripts. Therefore, the elements involved in spatial and temporal regulation of the xnf7 gene during development were contained within 421 basepairs upstream of the transcriptional initiation site. Future work will further define the elements involved in the spatial and temporal regulation and the trans-factors that interact with them. ^

Development of dendritic tonic GABAergic inhibition regulates excitability and plasticity in CA1 pyramidal neurons

Synaptic plasticity rules change during development: while hippocampal synapses can be potentiated by a single action potential pairing protocol in young neurons, mature neurons require burst firing to induce synaptic potentiation. An essential component for spike timing-dependent plasticity is the backpropagating action potential (BAP). BAP along the dendrites can be modulated by morphology and ion channel composition, both of which change during late postnatal development. However it is unclear whether these dendritic changes can explain the developmental changes in synaptic plasticity induction rules. Here, we show that tonic GABAergic inhibition regulates dendritic action potential backpropagation in adolescent but not pre-adolescent CA1 pyramidal neurons. These developmental changes in tonic inhibition also altered the induction threshold for spike timing-dependent plasticity in adolescent neurons. This GABAergic regulatory effect upon backpropagation is restricted to distal regions of apical dendrites (>200 μm) and mediated by α5-containing GABA(A) receptors. Direct dendritic recordings demonstrate α5-mediated tonic GABA(A) currents in adolescent neurons which can modulate backpropagating action potentials. These developmental modulations in dendritic excitability could not be explained by concurrent changes in dendritic morphology. To explain our data, model simulations propose a distally-increasing or localized distal expression of dendritic α5 tonic inhibition in mature neurons. Overall, our results demonstrate that dendritic integration and plasticity in more mature dendrites are significantly altered by tonic α5 inhibition in a dendritic region-specific and developmentally-regulated manner.

Age of the Peach Springs Tuff, Southeastern California and Western Arizona

Sanidine separates from pumice of the early Miocene Peach Springs Tuff are concordantly dated at 18.5 ± 0.2 Ma by two isotopic techniques. The Peach Springs Tuff is the only known unit that can be correlated between isolated outcrops of Miocene strata from the central Mojave Desert of southeastern California to the western Colorado Plateau in Arizona, across five structural provinces, a distance of 350 km. Thus the age of the Peach Springs Tuff is important to structural and paleogeographic reconstructions of a large region. Biotite and sanidine separates from bulk samples of the Peach Springs Tuff from zones of welding and vapor-phase alteration have not produced consistent ages by the K-Ar method. Published ages of mineral separates from 17 localities ranged from 16.2 to 20.5 Ma. Discordant 40Ar/39Ar incremental release spectra were obtained for one biotite and two of the sanidine separates. Ages that correspond to the last gas increments are as old as 27 Ma. The 40Ar/39Ar incremental release determinations on sanidine separated from blocks of Peach Springs Tuff pumice yield ages of 18.3 ± 0.3 and 18.6 ± 0.4 Ma. Laser fusion measurements yield a mean age of 18.51 ± 0.10. The results suggest that sanidine and biotite K-Ar ages older than about 18.5 Ma are due to inherited Ar from pre-Tertiary contaminants, which likely were incorporated into the tuff during deposition. Sanidine K-Ar ages younger than 18 Ma probably indicate incomplete extraction of radiogenic 40Ar, whereas laser fusion dates of biotite and hornblende younger than 18 Ma likely are due to postdepositional alteration. Laser fusion ages as high as 19.01 Ma on biotite grains from pumice suggest that minerals from pre-Tertiary country rocks also were incorporated in the magma chamber.

Climate change in Switzerland: a review of physical, institutional, and political aspects

Climate change is clearly discernible in observed climate records in Switzerland. It impacts on natural systems, ecosystems, and economic sectors such as agriculture, tourism, and energy, and it affects Swiss livelihood in various ways. The observed and projected changes call for a response from the political system, which in Switzerland is characterized by federalism and direct democratic instruments. Swiss climate science embraces natural and social sciences and builds on institutionalized links between researchers, public, and private stakeholders. In this article, we review the physical, institutional, and political aspects of climate change in Switzerland. We show how the current state of Swiss climate science and policy developed over the past 20 years in the context of international developments and national responses. Specific to Switzerland is its topographic setting with mountain regions and lowlands on both sides of the Alpine ridge, which makes climate change clearly apparent and for some aspects (tourist sector, hydropower, and extreme events) highly relevant and better perceivable (e.g., retreating glaciers). Not surprisingly the Alpine region is of central interest in Swiss climate change studies.

Organization of the inferotemporal cortex in the macaque monkey: Connections of areas PITv and CITvp

Visual cortex of macaque monkeys consists of a large number of cortical areas that span the occipital, parietal, temporal, and frontal lobes and occupy more than half of cortical surface. Although considerable progress has been made in understanding the contributions of many occipital areas to visual perceptual processing, much less is known concerning the specific functional contributions of higher areas in the temporal and frontal lobes. Previous behavioral and electrophysiological investigations have demonstrated that the inferotemporal cortex (IT) is essential to the animal's ability to recognize and remember visual objects. While it is generally recognized that IT consists of a number of anatomically and functionally distinct visual-processing areas, there remains considerable controversy concerning the precise number, size, and location of these areas. Therefore, the precise delineation of the cortical subdivisions of inferotemporal cortex is critical for any significant progress in the understanding of the specific contributions of inferotemporal areas to visual processing. In this study, anterograde and/or retrograde neuroanatomical tracers were injected into two visual areas in the ventral posterior and central portions of IT (areas PITv and CITvp) to elucidate the corticocortical connections of these areas with well known areas of occipital cortex and with less well understood regions of inferotemporal cortex. The locations of injection sites and the delineation of the borders of many occipital areas were aided by the pattern of interhemispheric connections, revealed following callosal transection and subsequent labeling with HRP. The resultant patterns of connections were represented on two-dimensional computational (CARET) and manual cortical maps and the laminar characteristics and density of the projection fields were quantified. The laminar and density features of these corticocortical connections demonstrate thirteen anatomically distinct subdivisions or areas distributed within the superior temporal sulcus and across the inferotemporal gyrus. These results serve to refine previous descriptions of inferotemporal areas, validate recently identified areas, and provide a new description of the hierarchical relationships among occipitotemporal cortical areas in macaques. ^

Systematic interpretation of molecular beacon polymerase chain reaction for identifying rpoB mutations in Mycobacterium tuberculosis isolates with mixed resistant and susceptible bacteria

Detection of multidrug-resistant tuberculosis (MDR-TB), a frequent cause of treatment failure, takes 2 or more weeks to identify by culture. RIF-resistance is a hallmark of MDR-TB, and detection of mutations in the rpoB gene of Mycobacterium tuberculosis using molecular beacon probes with real-time quantitative polymerase chain reaction (qPCR) is a novel approach that takes ≤2 days. However, qPCR identification of resistant isolates, particularly for isolates with mixed RIF-susceptible and RIF-resistant bacteria, is reader dependent and limits its clinical use. The aim of this study was to develop an objective, reader-independent method to define rpoB mutants using beacon qPCR. This would facilitate the transition from a research protocol to the clinical setting, where high-throughput methods with objective interpretation are required. For this, DNAs from 107 M. tuberculosis clinical isolates with known susceptibility to RIF by culture-based methods were obtained from 2 regions where isolates have not previously been subjected to evaluation using molecular beacon qPCR: the Texas–Mexico border and Colombia. Using coded DNA specimens, mutations within an 81-bp hot spot region of rpoB were established by qPCR with 5 beacons spanning this region. Visual and mathematical approaches were used to establish whether the qPCR cycle threshold of the experimental isolate was significantly higher (mutant) compared to a reference wild-type isolate. Visual classification of the beacon qPCR required reader training for strains with a mixture of RIF-susceptible and RIF-resistant bacteria. Only then had the visual interpretation by an experienced reader had 100% sensitivity and 94.6% specificity versus RIF-resistance by culture phenotype and 98.1% sensitivity and 100% specificity versus mutations based on DNA sequence. The mathematical approach was 98% sensitive and 94.5% specific versus culture and 96.2% sensitive and 100% specific versus DNA sequence. Our findings indicate the mathematical approach has advantages over the visual reading, in that it uses a Microsoft Excel template to eliminate reader bias or inexperience, and allows objective interpretation from high-throughput analyses even in the presence of a mixture of RIF-resistant and RIF-susceptible isolates without the need for reader training.^

Cross -packing among retroviruses and characterization of the feline immunodeficiency virus (FIV) packaging signal: Implications for the development of FIV-based gene transfer systems

Feline immunodeficiency virus (FIV)-based gene transfer systems are being seriously considered for human gene therapy as an alternative to vectors based on primate lentiviruses, a genetically complex group of retroviruses capable of infecting non-dividing cells. The greater phylogenetic distance between the feline and primate lentiviruses is thought to reduce chances of the generation of recombinant viruses. However, safety of FIV-based vector systems has not been tested experimentally. Since primate lentiviruses such as human and simian immunodeficiency viruses (HIV/SIV) can cross-package each other's genomes, we tested this trait with respect to FIV. Unexpectedly, both feline and primate lentiviruses were reciprocally able to both cross-package and propagate each other's RNA genomes. This was largely due to the recognition of viral packaging signals by the heterologous proteins. However, a simple retrovirus such as Mason-Pfizer monkey virus (MPMV) was unable to package FIV RNA. Interestingly, FIV could package MPMV RNA, but not propagate it for further steps of replication. These findings suggest that upon co-infection of the same host, cross-packaging may allow distinct retroviruses to generate chimeric variants with unknown pathogenic potential. ^ In order to understand the packaging determinants in FIV, we conducted a detailed mutational analysis of the region thought to contain FIV packaging signal. We show that the first 90–120 nt of the 5′ untranslated region (UTR) and the first 90 nt of gag were simultaneously required for efficient FIV RNA packaging. These results suggest that the primary FIV packaging signal is multipartite and discontinuous, composed of two core elements separated by 150 nt of the 5 ′UTR. ^ The above studies are being used towards the development of safer FIV-based self-inactivating (SIN) vectors. These vectors are being designed to eliminate the ability of FIV transfer vector RNAs to be mobilized by primate lentiviral proteins that may be present in the target cells. Preliminary test of the first generation of these vectors has revealed that they are incapable of being propagated by feline proteins. The inability of FIV transfer vectors to express packageable vector RNA after integration should greatly increase the safety of FIV vectors for human gene therapy. ^

Ice and snow measurements during Nathaniel B. Palmer cruise NBP0709 (SIMBA experiment)

The Sea Ice Mass Balance in the Antarctic (SIMBA) experiment was conducted from the RVIB N.B. Palmer in September and October 2007 in the Bellingshausen Sea in an area recently experiencing considerable changes in both climate and sea ice cover. Snow and ice properties were observed at 3 short-term stations and a 27-day drift station (Ice Station Belgica, ISB) during the winter-spring transition. Repeat measurements were performed on sea ice and snow cover at 5 ISB sites, each having different physical characteristics, with mean ice (snow) thicknesses varying from 0.6 m (0.1 m) to 2.3 m (0.7 m). Ice cores retrieved every five days from 2 sites and measured for physical, biological, and chemical properties. Three ice mass-balance buoys (IMBs) provided continuous records of snow and ice thickness and temperature. Meteorological conditions changed from warm fronts with high winds and precipitation followed by cold and calm periods through four cycles during ISB. The snow cover regulated temperature flux and controlled the physical regime in which sea ice morphology changed. Level thin ice areas had little snow accumulation and experienced greater thermal fluctuations resulting in brine salinity and volume changes, and winter maximum thermodynamic growth of ~0.6 m in this region. Flooding and snow-ice formation occurred during cold spells in ice and snow of intermediate thickness. In contrast, little snow-ice formed in flooded areas with thicker ice and snow cover, instead nearly isothermal, highly permeable ice persisted. In spring, short-lived cold air episodes did not effectively penetrate the sea ice nor overcome the effect of ocean heat flux, thus favoring net ice thinning from bottom melt over ice thickening from snow-ice growth, in all cases. These warm ice conditions were consistent with regional remote sensing observations of earlier ice breakup and a shorter sea ice season, more recently observed in the Bellingshausen Sea.

Living benthic foraminifers in bottom sediments of the southeastern Kandalaksha Bay (White Sea)

Analysis of composition and distribution of benthic foraminifers in six samples of bottom sediments obtained in the southeast Kandalaksha Bay of the White Sea at water depths of 20 to 155 m revealed their dependence on lithology and different hydrological characteristics. It is shown that living foraminifers populating relatively shallow areas of the bay (20-60 m), which are bathed by seasonally warmed intermediate water with temperature 0.7-1.5°C and salinity 26 per mil, are characterized by high abundance (250-750 specimens/10 ccm of wet sediment) and prevalence of agglutinated species (Eggerella advena, Recurvoides turbinatus, and others). Deeper (155 m) where cold and relatively saline deep water occurs (-1.4°C, 29.5 per mil), abundance is an order lower (30 specimens/10 ccm) and is dominated by calcareous taxa Cassidulina reniforme, an Arctic cold resistant species.

Distribution of Recent benthic foraminifera in Arctic galciomarine sediments

Foraminifera were examined in recent (<100 years) fine-grained glaciomarine muds from surface sediments and cores from Nordensheld Bay, Novaja Zemlja, and Hornsund and Bellsund, Spitsbergen. This study presents the first data on modern foraminifera distribution for fjord environments in Novaja Zemlja, Russia. The data are interpreted with reference to the distribution of foraminiferal near Svalbard and the Barents Sea. In Nordensheld Bay, live and dead Nonionellina labradorica and Islandiella norcrossi are most abundant in the outer fjord. Cassidulina reniforme and Allogromiina spp. dominate in the middle and inner fjord. The dominant species are dissimilar to species occurring in other areas of the Barents Sea region, with the exception of Svalbard fjords. The number of live foraminifera (24 to 122 tests/10 cm1) in outer and middle Nordensheld Bay corresponds with values known from the open Barents Sea. However, the biomass (0.03 mg/10 cm**3) is two orders of magnitude less due to smaller foraminiferal test size, which in glaciomarine sediments reflects the absence of larger species, paucity of large specimens, and high occurrence of juvenile foraminifera. The smaller size indicates an opportunistic response to environmental stress due to glacier proximity. The presence of Quinqueloculina stalkeri is diagnostic of glaciomarine environments in fjords of Novaja Zemlja and Svalbard.

Análisis de co-términos y de redes sociales para la generación de mapas temáticos

La mejora de las capacidades de búsqueda y de las interfaces de los opacs sigue siendo uno de los principales desafíos para las bibliotecas, especialmente en lo que respecta al acceso por materias. Las interfaces visuales pueden facilitar la recuperación. El objetivo del presente trabajo es explorar si la combinación de técnicas de análisis de co-términos y de redes sociales resulta ser una metodología válida para la generación de mapas temáticos de la colección. La principal conclusión es que el método es válido, y que los mapas obtenidos podrían servir como interfaz visual para el acceso por materias. También es útil para detectar problemas en los criterios de indización y contribuir a la mejora de la calidad de la descripción temática del conjunto documental

High-resolution ice thickness and bed topography of a land-terminating section of the Greenland Ice Sheet, with links to GeoTIFFs and text files

We present ice thickness and bed topography maps with a high spatial resolution (250-500 m) of a land-terminating section of the Greenland Ice Sheet derived from ground-based and airborne radar surveys. The data have a total area of ~12 000 km^2 and cover the whole ablation area of the outlet glaciers of Isunnguata Sermia, Russell, Leverett, Ørkendalen and Isorlersuup up to the long-term mass balance equilibrium line altitude at ~1600 m above sea level. The bed topography shows highly variable subglacial trough systems, and the trough of Isunnguata Sermia Glacier is overdeepened and reaches an elevation of ~500 m below sea level. The ice surface is smooth and only reflects the bedrock topography in a subtle way, resulting in a highly variable ice thickness. The southern part of our study area consists of higher bed elevations compared to the northern part. The compiled data sets of ground-based and airborne radar surveys cover one of the most studied regions of the Greenland Ice Sheet and can be valuable for detailed studies of ice sheet dynamics and hydrology.

Análisis de co-términos y de redes sociales para la generación de mapas temáticos

La mejora de las capacidades de búsqueda y de las interfaces de los opacs sigue siendo uno de los principales desafíos para las bibliotecas, especialmente en lo que respecta al acceso por materias. Las interfaces visuales pueden facilitar la recuperación. El objetivo del presente trabajo es explorar si la combinación de técnicas de análisis de co-términos y de redes sociales resulta ser una metodología válida para la generación de mapas temáticos de la colección. La principal conclusión es que el método es válido, y que los mapas obtenidos podrían servir como interfaz visual para el acceso por materias. También es útil para detectar problemas en los criterios de indización y contribuir a la mejora de la calidad de la descripción temática del conjunto documental

Análisis de co-términos y de redes sociales para la generación de mapas temáticos

La mejora de las capacidades de búsqueda y de las interfaces de los opacs sigue siendo uno de los principales desafíos para las bibliotecas, especialmente en lo que respecta al acceso por materias. Las interfaces visuales pueden facilitar la recuperación. El objetivo del presente trabajo es explorar si la combinación de técnicas de análisis de co-términos y de redes sociales resulta ser una metodología válida para la generación de mapas temáticos de la colección. La principal conclusión es que el método es válido, y que los mapas obtenidos podrían servir como interfaz visual para el acceso por materias. También es útil para detectar problemas en los criterios de indización y contribuir a la mejora de la calidad de la descripción temática del conjunto documental

Physical properties and elastic constants of ODP Holes 137-504B and 140-504B upper crustal rocks

Seismic velocities have been measured at confining pressures of 100 MPa and 600 MPa for sheeted dike samples recovered during Ocean Drilling Program Legs 137 and 140. The compressional- and shear-wave velocities show an increase with depth at Hole 504B, which is in sharp contrast to the atmospheric pressure velocity measurements performed as part of the shipboard analyses. Rocks exposed to different types of alteration and fracture patterns show distinct changes in their physical properties. The seismic reflectors observed on the vertical seismic profile (VSP) experiment performed during Leg 111 may have been caused by low velocity zones resulting from alteration. The amount of fracturing and hydrothermal alteration in several zones also may have contributed to the acoustic impedance contrast necessary to produce the E5 reflector. Poisson's ratios calculated from laboratory velocity measurements show several low values at depths ranging from 1600 mbsf to 2000 mbsf, which tends to follow similar trends obtained from previous oceanic refraction experiments. A comparison of physical properties between samples recovered from Hole 504B and ophiolite studies in the Bay of Islands and Oman shows a good correlation with the Bay of Islands but significant differences from the measurements performed in the Oman complex.