Detection of competing DNA structures by thermal gradient gel electrophoresis: from self-association to triple helix formation by (G,A)-containing oligonucleotides

Sequence-specific recognition of DNA can be achieved by triple helix-forming oligonucleotides that bind to the major groove of double-helical DNA. These oligonucleotides have been used as sequence-specific DNA ligands for various purposes, including sequence-specific gene regulation in the so-called ‘antigene strategy’. In particular, (G,A)-containing oligonucleotides can form stable triple helices under physiological conditions. However, triplex formation may be in competition with self-association of these oligonucleotides. For biological applications it would be interesting to identify the conditions under which one structure is favoured as compared to the other(s). Here we have directly studied competition between formation of a parallel (G,A) homoduplex and that of a triple helix by a 13 nt (G,A)-containing oligonucleotide. Temperature gradient gel electrophoresis allows simultaneous detection of competition between the two structures, because of their different temperature dependencies and gel electrophoretic mobilities, and characterisation of this competition.

Saline Stress Alters the Temporal Patterns of Xylem Differentiation and Alternative Oxidase Expression in Developing Soybean Roots1

We conducted a coordinated biochemical and morphometric analysis of the effect of saline conditions on the differentiation zone of developing soybean (Glycine max L.) roots. Between d 3 and d 14 for seedlings grown in control or NaCl-supplemented medium, we studied (a) the temporal evolution of the respiratory alternative oxidase (AOX) capacity in correlation with the expression and localization of AOX protein analyzed by tissue-print immunoblotting; (b) the temporal evolution and tissue localization of a peroxidase activity involved in lignification; and (c) the structural changes, visualized by light microscopy and quantified by image digitization. The results revealed that saline stress retards primary xylem differentiation. There is a corresponding delay in the temporal pattern of AOX expression, which is consistent with the xylem-specific localization of AOX protein and the idea that this enzyme is linked to xylem development. An NaCl-induced acceleration of the development of secondary xylem was also observed. However, the temporal pattern of a peroxidase activity localized in the primary and secondary xylem was unaltered by NaCl treatment. Thus, the NaCl-stressed root was specifically affected in the temporal patterns of AOX expression and xylem development.

An experimental system for analyzing response to a morphogen gradient.

A recently described experimental system for analyzing the mode of action of a morphogen gradient involves the in situ hybridization of sectioned tissue constructs. In these constructs, a source of activin signaling induces the transcription of several mesodermal genes in blastula animal caps, according to the position of cells in a concentration gradient. New experiments show that activin-loaded beads emit a signal for only 2 hr and that the same cell can be induced to express different genes. We determine the position in the gradient and the time after the start of activin signaling at which early genes, including Mix1, Xpo, Xwnt8, Xchd, and Xlim1, are activated, relative to the previously tested genes Xbra and Xgsc.

prlA suppressors in Escherichia coli relieve the proton electrochemical gradient dependency of translocation of wild-type precursors.

The SecY protein of Escherichia coli is an integral membrane component of the protein export apparatus. Suppressor mutations in the secY gene (prlA alleles) have been isolated that restore the secretion of precursor proteins with defective signal sequences. These mutations have never been shown to affect the translocation of wild-type precursor proteins. Here, we report that prlA suppressor mutations relieve the proton-motive force (pmf) dependency of the translocation of wild-type precursors, both in vivo and in vitro. Furthermore, the proton-motive force dependency of the translocation of a precursor with a stably folded domain in the mature region was suppressed by prlA mutations in vitro. These data show that prlA mutations cause a general relaxation of the export apparatus rather than a specific change that results in bypassing of the recognition of the signal sequence. In addition, these results are indicative for a mechanism in which the proton-motive force stimulates translocation by altering the conformation of the translocon.

Relationship between photosynthetic electron transport and pH gradient across the thylakoid membrane in intact leaves.

Under conditions (0.2% CO2; 1% O2) that allow high rates of photosynthesis, chlorophyll fluorescence was measured simultaneously with carbon assimilation at various light intensities in spinach (Spinacia oleracea) leaves. Using a stoichiometry of 3 ATP/CO2 and the known relationship between ATP synthesis rate and driving force (Delta pH), we calculated the light-dependent pH gradient (Delta pH) across the thylakoid membrane in intact leaves. These Delta pH values were correlated with the photochemical (qP) and nonphotochemical (qN) quenching of chlorophyll fluorescence and with the quantum yield of photosystem II (phiPSII). At Delta pH > 2.1 all three parameters (qP, qN, and phiPSII) changed very steeply with increasing DeltapH (decreasing pH in the thylakoid). The observed pH dependences followed hexacooperative titration curves with slightly different pKa values. The significance of the steep pH dependences with slightly different pKa values is discussed in relation to the regulation of photosynthetic electron transport in intact leaves.

A gradient estimate to a degenerate parabolic equation with a singular absorption term: The global quenching phenomena

We prove global existence of nonnegative solutions to the one dimensional degenerate parabolic problems containing a singular term. We also show the global quenching phenomena for L1 initial datums. Moreover, the free boundary problem is considered in this paper.

Remote Sensing of Vegetation Change Across a Latitudinal Gradient in the Canadian Arctic

Global air surface temperatures and precipitation have increased over the last several decades resulting in a trend of greening across the Circumpolar Arctic. The spatial variability of warming and the inherent effects on plant communities has not proven to be uniform or homogeneous on global or local scales. We can apply remote sensing vegetation indices such as the Normalized Difference Vegetation Index (NDVI) to map and monitor vegetation change (e.g., phenology, greening, percent cover, and biomass) over time. It is important to document how Arctic vegetation is changing, as it will have large implications related to global carbon and surface energy budgets. The research reported here examined vegetation greening across different spatial and temporal scales at two disparate Arctic sites: Apex River Watershed (ARW), Baffin Island, and Cape Bounty Arctic Watershed Observatory (CBAWO), Melville Island, NU. To characterize the vegetation in the ARW, high spatial resolution WorldView-2 data were processed to create a supervised land-cover classification and model percent vegetation cover (PVC) (a similar process had been completed in a previous study for the CBAWO). Meanwhile, NDVI data spanning the past 30 years were derived from intermediate resolution Landsat data at the two Arctic sites. The land-cover classifications at both sites were used to examine the Landsat NDVI time series by vegetation class. Climate variables (i.e., temperature, precipitation and growing season length (GSL) were examined to explore the potential relationships of NDVI to climate warming. PVC was successfully modeled using high resolution data in the ARW. PVC and plant communities appear to reside along a moisture and altitudinal gradient. The NDVI time series demonstrated an overall significant increase in greening at the CBAWO (High Arctic site), specifically in the dry and mesic vegetation type. However, similar overall greening was not observed for the ARW (Low Arctic site). The overall increase in NDVI at the CBAWO was attributed to a significant increase in July temperatures, precipitation and GSL.

A randomized trial of nebulized 3% hypertonic saline with salbutamol in the treatment of acute bronchiolitis in hospitalized infants

OBJECTIVE: Acute bronchiolitis is a common disorder of infants that often results in hospitalization. Apart from supportive care, no therapy has been shown to influence the course of the disease, except for a possible effect of nebulized hypertonic saline (HS). To determine whether this does have beneficial effects on length of stay in hospital or on severity scores, we undertook a double-blind, randomized, controlled trial in a pediatric department of a Portuguese hospital. METHODS: Previously healthy infants, younger than 12 months, hospitalized with mild-to-moderate acute viral bronchiolitis were randomized to receive either nebulized 3% (hypertonic, HS) or 0.9% (normal, NS) saline during their entire hospital stay. Primary endpoints were: length of hospital stay and severity scores on each day of hospitalization. Need for supplemental oxygen, further add-on medications and adverse effects were also analyzed. RESULTS: Sixty-eight patients completed the study (HS: 33; NS: 35). The median length of hospital stay did not differ between groups: HS: 5.6 ± 2.3 days; NS: 5.4 ± 2.1 days (P = 0.747). We found no difference between groups in severity scores from day 1 to day 4. There were no differences in need for supplemental oxygen or add-on medications. Patients in HS group had significantly more cough (46% vs. 20%, P = 0.025) and rhinorrhoe (58% vs. 31%, P = 0.30). CONCLUSION: This study does not support the use of nebulized HS over NS in therapy of hospitalized children with mild-to-moderate acute viral bronchiolitis

Gradient-Domain Bidirectional Path Tracing

Gradient-domain path tracing has recently been introduced as an efficient realistic image synthesis algorithm. This paper introduces a bidirectional gradient-domain sampler that outperforms traditional bidirectional path tracing often by a factor of two to five in terms of squared error at equal render time. It also improves over unidirectional gradient-domain path tracing in challenging visibility conditions, similarly as conventional bidirectional path tracing improves over its unidirectional counterpart. Our algorithm leverages a novel multiple importance sampling technique and an efficient implementation of a high-quality shift mapping suitable for bidirectional path tracing. We demonstrate the versatility of our approach in several challenging light transport scenarios.

Plant species, biomass and environmental characteristics of relevés along the North America Arctic bioclimate gradient

Question: How do interactions between the physical environment and biotic properties of vegetation influence the formation of small patterned-ground features along the Arctic bioclimate gradient? Location: At 68° to 78°N: six locations along the Dalton Highway in arctic Alaska and three in Canada (Banks Island, Prince Patrick Island and Ellef Ringnes Island). Methods: We analysed floristic and structural vegetation, biomass and abiotic data (soil chemical and physical parameters, the n-factor [a soil thermal index] and spectral information [NDVI, LAI]) on 147 microhabitat releves of zonalpatterned-ground features. Using mapping, table analysis (JUICE) and ordination techniques (NMDS). Results: Table analysis using JUICE and the phi-coefficient to identify diagnostic species revealed clear groups of diagnostic plant taxa in four of the five zonal vegetation complexes. Plant communities and zonal complexes were generally well separated in the NMDS ordination. The Alaska and Canada communities were spatially separated in the ordination because of different glacial histories and location in separate floristic provinces, but there was no single controlling environmental gradient. Vegetation structure, particularly that of bryophytes and total biomass, strongly affected thermal properties of the soils. Patterned-ground complexes with the largest thermal differential between the patterned-ground features and the surrounding vegetation exhibited the clearest patterned-ground morphologies.

Pollen abundance in Late Quaternary sediment cores off West Africa

The pollen record of three marine late Quaternary cores off Senegal shows a juxtaposition of Mediterranean, Northern Saharan, Central Saharan elements, which are considered transported by the trade winds from a winter-rainfall area, and Sahelian, Soudanese, Soudano-Guinean elements, considered transported both by winds and mostly by the Senegal River, and coming from the monsoonal, summer tropical rainfall area of southern West Africa. Littoral vegetation is either the edaphically dry and saline Chenopodiaceae from sebkhas at the time of the main regression, or the warm tropical humid mangrove with Rhizophora during the humid optimum period. Four stratigraphic zones reflect, from basis to top: Zone 4. A semi-arid period with a balanced pollen input. Zone 3. A very arid period with the disappearance of monsoonal pollen, probably from the disappearance of the Senegal River, a very saline littoral plain with Chenopodiaceae, a larger input of northern Saharan pollen from intensified trade winds. Zone 2. A quite humid period, much more so than today, very suddenly established, with a northward extension of the monsoonal areas, a rich littoral mangrove, and weakening of the trade winds. Zone l. A slow and steady evolution toward the present semi-humid conditions with regression of the mangrove, and of the monsoonal areas toward the south. Tentative datations and correlations with the Tchad area suggested: zone 4: 22,500 to 19,000 years BP; zone 3: 19,000 to 12,500 years BP; zone 2: 12,500 to 5,500 years BP; zone 1: 5,500 years BP to top of core. Dinoflagellate cysts display a tropical assemblage with mostly estuarine neritic elements and also a weak oceanic component, mostly in the lower slope core 47. Cosmopolitan taxa dominate the assemblage and only a few species point to more specialized environments. Quantitative variations of the assemblage are the basis of stratigraphy which is not similar to the pollen stratigraphy, and an inshore-outshore gradient has to be taken into account to correlate the three cores.

Saline Township (Mich.) residences and business, ca. 1874

Left to right: Res. of Orange Risdon, Sec. 1, Saline Tp. Mich.; Res. of Andrew Bush, Sec. 3, Saline Tp. Mich.; Res. of Dudley Miller, Sec. 24, Saline Tp. Mich.; Saline Exchange, A. Harmon Prop, Saline, Mich.; Res. of Abram Davenport, Sec. 36, Saline Tp. Mich.; Res. of Milton Reynolds, Sec. 22, Saline Tp. Mich.; Res. of George Neissle, Sec. 12, Saline Tp. Mich.; Res. of Robert Hammond, Sec. 25, Saline Tp. Mich.; Res. of M. Himerdinger, Sec. 8, Saline Tp. Mich. Publication information: Chicago, Ill. : Everts & Stewart, 1874.

Map of Saline Township. T.P. No. 4 S. Range No. 5 E [plat]

Publication information: Chicago, Ill. : Everts & Stewart, 1874.