2,864 research outputs found
Using zeta-potential measurements to quantify peptide partition to lipid membranes
© The Author(s) 2011. This article is published with open access at Springerlink.com.Open Access: This article is distributed under the terms of the Creative Commons Attribution Noncommercial License which permits any noncommercial use, distribution, and reproduction in any medium, provided the original author(s) and source are credited.Many cellular phenomena occur on the biomembranes. There are plenty of molecules (natural or xenobiotics) that interact directly or partially with the cell membrane. Biomolecules, such as several peptides (e.g., antimicrobial peptides) and proteins, exert their effects at the cell membrane level. This feature makes necessary investigating their interactions with lipids to clarify their mechanisms of action and side effects necessary. The determination of molecular lipid/water partition constants (Kp) is frequently used to quantify the extension of the interaction. The determination of this parameter has been achieved by using different methodologies, such as UV-Vis absorption spectrophotometry, fluorescence spectroscopy and ζ-potential measurements. In this work, we derived and tested a mathematical model to determine the Kp from ζ-potential data. The values obtained with this method were compared with those obtained by fluorescence spectroscopy, which is a regular technique used to quantify the interaction of intrinsically fluorescent peptides with selected biomembrane model systems. Two antimicrobial peptides (BP100 and pepR) were evaluated by this new method. The results obtained by this new methodology show that ζ-potential is a powerful technique to quantify peptide/lipid interactions of a wide variety of charged molecules, overcoming some of the limitations inherent to other techniques, such as the need for fluorescent labeling.This work was partially supported by project PTDC/QUI/ 69937/2006 from Fundação para a Ciência e Tecnologia-Ministério da Ciência, Tecnologia e Ensino Superior (FCT-MCTES, Portugal), and by Fundação Calouste Gulbenkian (Portugal). JMF and MMD also thank FCT-MCTES for grants IMM/BT/37-2010 and SFRH/BD/41750/2007, respectively
Fructose transport-deficient Staphylococcus aureus reveals important role of epithelial glucose transporters in limiting sugar-driven bacterial growth in airway surface liquid.
Hyperglycaemia as a result of diabetes mellitus or acute illness is associated with increased susceptibility to respiratory infection with Staphylococcus aureus. Hyperglycaemia increases the concentration of glucose in airway surface liquid (ASL) and promotes the growth of S. aureus in vitro and in vivo. Whether elevation of other sugars in the blood, such as fructose, also results in increased concentrations in ASL is unknown and whether sugars in ASL are directly utilised by S. aureus for growth has not been investigated. We obtained mutant S. aureus JE2 strains with transposon disrupted sugar transport genes. NE768(fruA) exhibited restricted growth in 10 mM fructose. In H441 airway epithelial-bacterial co-culture, elevation of basolateral sugar concentration (5-20 mM) increased the apical growth of JE2. However, sugar-induced growth of NE768(fruA) was significantly less when basolateral fructose rather than glucose was elevated. This is the first experimental evidence to show that S. aureus directly utilises sugars present in the ASL for growth. Interestingly, JE2 growth was promoted less by glucose than fructose. Net transepithelial flux of D-glucose was lower than D-fructose. However, uptake of D-glucose was higher than D-fructose across both apical and basolateral membranes consistent with the presence of GLUT1/10 in the airway epithelium. Therefore, we propose that the preferential uptake of glucose (compared to fructose) limits its accumulation in ASL. Pre-treatment with metformin increased transepithelial resistance and reduced the sugar-dependent growth of S. aureus. Thus, epithelial paracellular permeability and glucose transport mechanisms are vital to maintain low glucose concentration in ASL and limit bacterial nutrient sources as a defence against infection
Belowground DNA-based techniques: untangling the network of plant root interactions
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Urinary Proteomics to Support Diagnosis of Stroke
Accurate diagnosis in suspected ischaemic stroke can be difficult. We explored the urinary proteome in patients with stroke (n = 69), compared to controls (n = 33), and developed a biomarker model for the diagnosis of stroke. We performed capillary electrophoresis online coupled to micro-time-of-flight mass spectrometry. Potentially disease-specific peptides were identified and a classifier based on these was generated using support vector machine-based software. Candidate biomarkers were sequenced by liquid chromatography-tandem mass spectrometry. We developed two biomarker-based classifiers, employing 14 biomarkers (nominal p-value <0.004) or 35 biomarkers (nominal p-value <0.01). When tested on a blinded test set of 47 independent samples, the classification factor was significantly different between groups; for the 35 biomarker model, median value of the classifier was 0.49 (−0.30 to 1.25) in cases compared to −1.04 (IQR −1.86 to −0.09) in controls, p<0.001. The 35 biomarker classifier gave sensitivity of 56%, specificity was 93% and the AUC on ROC analysis was 0.86. This study supports the potential for urinary proteomic biomarker models to assist with the diagnosis of acute stroke in those with mild symptoms. We now plan to refine further and explore the clinical utility of such a test in large prospective clinical trials
Water relations of evergreen and drought-deciduous trees along a seasonally dry tropical forest chronosequence
Seasonally dry tropical forests (SDTF) are characterized by pronounced seasonality in rainfall, and as a result trees in these forests must endure seasonal variation in soil water availability. Furthermore, SDTF on the northern Yucatan Peninsula, Mexico, have a legacy of disturbances, thereby creating a patchy mosaic of different seral stages undergoing secondary succession. We examined the water status of six canopy tree species, representing contrasting leaf phenology (evergreen vs. drought-deciduous) at three seral stages along a fire chronosequence in order to better understand strategies that trees use to overcome seasonal water limitations. The early-seral forest was characterized by high soil water evaporation and low soil moisture, and consequently early-seral trees exhibited lower midday bulk leaf water potentials (ΨL) relative to late-seral trees (−1.01 ± 0.14 and −0.54 ± 0.07 MPa, respectively). Although ΨL did not differ between evergreen and drought-deciduous trees, results from stable isotope analyses indicated different strategies to overcome seasonal water limitations. Differences were especially pronounced in the early-seral stage where evergreen trees had significantly lower xylem water δ18O values relative to drought-deciduous trees (−2.6 ± 0.5 and 0.3 ± 0.6‰, respectively), indicating evergreen species used deeper sources of water. In contrast, drought-deciduous trees showed greater enrichment of foliar 18O (∆18Ol) and 13C, suggesting lower stomatal conductance and greater water-use efficiency. Thus, the rapid development of deep roots appears to be an important strategy enabling evergreen species to overcome seasonal water limitation, whereas, in addition to losing a portion of their leaves, drought-deciduous trees minimize water loss from remaining leaves during the dry season
Physics Opportunities of e+e- Linear Colliders
We describe the anticipated experimental program of an e+e- linear collider
in the energy range 500 GeV -- 1.5 TeV. We begin with a description of current
collider designs and the expected experimental environment. We then discuss
precision studies of the W boson and top quark. Finally, we review the range of
models proposed to explain the physics of electroweak symmetry breaking and
show, for each case, the central role that the linear collider experiments will
play in elucidating this physics. (to appear in Annual Reviews of Nuclear and
Particle Science)Comment: 93 pages, latex + 23 figures; typos corrections + 1 reference adde
Seasonal variation of water uptake of a Quercus suber tree in Central Portugal
Hydraulic redistribution (HR) is the phenomenon
where plant roots transfer water between
soil horizons of different water potential. When dry
soil is a stronger sink for water loss from the plant
than transpiration, water absorbed by roots in wetter
soil horizons is transferred toward, and exuded into
dry soil via flow reversals through the roots. Reverse
flow is a good marker of HR and can serve as a useful
tool to study it over the long-term. Seasonal variation
of water uptake of a Quercus suber tree was studied
from late winter through autumn 2003 at Rio Frio
near Lisbon, Portugal. Sap flow was measured in five
small shallow roots (diameter of 3–4 cm), 1 to 2 m
from the tree trunk and in four azimuths and at
different xylem depths at the trunk base, using the
heat field deformation method (HFD). The pattern of
sap flow differed among lateral roots as soil dried with constant positive flow in three roots and reverse
flow in two other roots during the night when
transpiration ceased. Rain modified the pattern of
flow in these two roots by eliminating reverse flow
and substantially increasing water uptake for transpiration
during the day. The increase in water uptake in
three other roots following rain was not so substantial.
In addition, the flux in individual roots was correlated
to different degrees with the flux at different radial
depths and azimuthal directions in trunk xylem. The
flow in outer trunk xylem seemed to be mostly
consistent with water movement from surface soil
horizons, whereas deep roots seemed to supply water
to the whole cross-section of sapwood. When water
flow substantially decreased in shallow lateral roots
and the outer stem xylem during drought, water flow
in the inner sapwood was maintained, presumably due
to its direct connection to deep roots. Results also
suggest the importance of the sap flow sensor
placement, in relation to sinker roots, as to whether
lateral roots might be found to exhibit reverse flow
during drought. This study is consistent with the
dimorphic rooting habit of Quercus suber trees in
which deep roots access groundwater to supply
superficial roots and the whole tree, when shallow
soil layers were dry
Retinal repair by transplantation of photoreceptor precursors
Photoreceptor loss causes irreversible blindness in many retinal diseases. Repair of such damage by cell transplantation is one of the most feasible types of central nervous system repair; photoreceptor degeneration initially leaves the inner retinal circuitry intact and new photoreceptors need only make single, short synaptic connections to contribute to the retinotopic map. So far, brain- and retina-derived stem cells transplanted into adult retina have shown little evidence of being able to integrate into the outer nuclear layer and differentiate into new photoreceptors(1-4). Furthermore, there has been no demonstration that transplanted cells form functional synaptic connections with other neurons in the recipient retina or restore visual function. This might be because the mature mammalian retina lacks the ability to accept and incorporate stem cells or to promote photoreceptor differentiation. We hypothesized that committed progenitor or precursor cells at later ontogenetic stages might have a higher probability of success upon transplantation. Here we show that donor cells can integrate into the adult or degenerating retina if they are taken from the developing retina at a time coincident with the peak of rod genesis(5). These transplanted cells integrate, differentiate into rod photoreceptors, form synaptic connections and improve visual function. Furthermore, we use genetically tagged postmitotic rod precursors expressing the transcription factor Nrl (ref. 6) ( neural retina leucine zipper) to show that successfully integrated rod photoreceptors are derived only from immature post-mitotic rod precursors and not from proliferating progenitor or stem cells. These findings define the ontogenetic stage of donor cells for successful rod photoreceptor transplantation.Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/62596/1/nature05161.pd
Planetary Rings
Planetary rings are the only nearby astrophysical disks, and the only disks
that have been investigated by spacecraft. Although there are significant
differences between rings and other disks, chiefly the large planet/ring mass
ratio that greatly enhances the flatness of rings (aspect ratios as small as
1e-7), understanding of disks in general can be enhanced by understanding the
dynamical processes observed at close-range and in real-time in planetary
rings. We review the known ring systems of the four giant planets, as well as
the prospects for ring systems yet to be discovered. We then review planetary
rings by type. The main rings of Saturn comprise our system's only dense broad
disk and host many phenomena of general application to disks including spiral
waves, gap formation, self-gravity wakes, viscous overstability and normal
modes, impact clouds, and orbital evolution of embedded moons. Dense narrow
rings are the primary natural laboratory for understanding shepherding and
self-stability. Narrow dusty rings, likely generated by embedded source bodies,
are surprisingly found to sport azimuthally-confined arcs. Finally, every known
ring system includes a substantial component of diffuse dusty rings. Planetary
rings have shown themselves to be useful as detectors of planetary processes
around them, including the planetary magnetic field and interplanetary
impactors as well as the gravity of nearby perturbing moons. Experimental rings
science has made great progress in recent decades, especially numerical
simulations of self-gravity wakes and other processes but also laboratory
investigations of coefficient of restitution and spectroscopic ground truth.
The age of self-sustained ring systems is a matter of debate; formation
scenarios are most plausible in the context of the early solar system, while
signs of youthfulness indicate at least that rings have never been static
phenomena.Comment: 82 pages, 34 figures. Final revision of general review to be
published in "Planets, Stars and Stellar Systems", P. Kalas and L. French
(eds.), Springer (http://refworks.springer.com/sss
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