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71.
Colzani M Schütz F Potts A Waridel P Quadroni M 《Molecular & cellular proteomics : MCP》2008,7(5):927-937
Metabolic labeling techniques have recently become popular tools for the quantitative profiling of proteomes. Classical stable isotope labeling with amino acids in cell cultures (SILAC) uses pairs of heavy/light isotopic forms of amino acids to introduce predictable mass differences in protein samples to be compared. After proteolysis, pairs of cognate precursor peptides can be correlated, and their intensities can be used for mass spectrometry-based relative protein quantification. We present an alternative SILAC approach by which two cell cultures are grown in media containing isobaric forms of amino acids, labeled either with 13C on the carbonyl (C-1) carbon or 15N on backbone nitrogen. Labeled peptides from both samples have the same nominal mass and nearly identical MS/MS spectra but generate upon fragmentation distinct immonium ions separated by 1 amu. When labeled protein samples are mixed, the intensities of these immonium ions can be used for the relative quantification of the parent proteins. We validated the labeling of cellular proteins with valine, isoleucine, and leucine with coverage of 97% of all tryptic peptides. We improved the sensitivity for the detection of the quantification ions on a pulsing instrument by using a specific fast scan event. The analysis of a protein mixture with a known heavy/light ratio showed reliable quantification. Finally the application of the technique to the analysis of two melanoma cell lines yielded quantitative data consistent with those obtained by a classical two-dimensional DIGE analysis of the same samples. Our method combines the features of the SILAC technique with the advantages of isobaric labeling schemes like iTRAQ. We discuss advantages and disadvantages of isobaric SILAC with immonium ion splitting as well as possible ways to improve it. 相似文献
72.
Daniel H. Libraty Luz P. Acosta Veronica Tallo Edelwisa Segubre-Mercado Analisa Bautista James A. Potts Richard G. Jarman In-Kyu Yoon Robert V. Gibbons Job D. Brion Rosario Z. Capeding 《PLoS medicine》2009,6(10)
Background
Dengue hemorrhagic fever (DHF) is the severe and life-threatening syndrome that can develop after infection with any one of the four dengue virus (DENV) serotypes. DHF occurs almost exclusively in individuals with secondary heterologous DENV infections and infants with primary DENV infections born to dengue immune mothers. The widely accepted explanation for the pathogenesis of DHF in these settings, particularly during infancy, is antibody-dependent enhancement (ADE) of DENV infection.Methods and Findings
We conducted a prospective nested case-control study of DENV infections during infancy. Clinical data and blood samples were collected from 4,441 mothers and infants in up to two pre-illness study visits, and surveillance was performed for symptomatic and inapparent DENV infections. Pre-illness plasma samples were used to measure the associations between maternally derived anti-DENV3 antibody-neutralizing and -enhancing capacities at the time of DENV3 infection and development of infant DHF.The study captured 60 infants with DENV infections across a wide spectrum of disease severity. DENV3 was the predominant serotype among the infants with symptomatic (35/40) and inapparent (15/20) DENV infections, and 59/60 infants had a primary DENV infection. The estimated in vitro anti-DENV3 neutralizing capacity at birth positively correlated with the age of symptomatic primary DENV3 illness in infants. At the time of symptomatic DENV3 infection, essentially all infants had low anti-DENV3 neutralizing activity (50% plaque reduction neutralizing titers [PRNT50] ≤50) and measurable DENV3 ADE activity. The infants who developed DHF did not have significantly higher frequencies or levels of DENV3 ADE activity compared to symptomatic infants without DHF. A higher weight-for-age in the first 3 mo of life and at illness presentation was associated with a greater risk for DHF from a primary DENV infection during infancy.Conclusions
This prospective nested case-control study of primarily DENV3 infections during infancy has shown that infants exhibit a full range of disease severity after primary DENV infections. The results support an initial in vivo protective role for maternally derived antibody, and suggest that a DENV3 PRNT50 >50 is associated with protection from symptomatic DENV3 illness. We did not find a significant association between DENV3 ADE activity at illness onset and the development of DHF compared with less severe symptomatic illness. The results of this study should encourage rethinking or refinement of the current ADE pathogenesis model for infant DHF and stimulate new directions of research into mechanisms responsible for the development of DHF during infancy.Trial registration
ClinicalTrials.gov Please see later in the article for the Editors'' Summary NCT00377754相似文献73.
Sabitha Prabhakaran Xiaowen Liang Jonathan T. Skare Jennifer R. Potts Magnus H??k 《PloS one》2009,4(4)
Background
BBK32 is a surface expressed lipoprotein and fibronectin (Fn)-binding microbial surface component recognizing adhesive matrix molecule (MSCRAMM) of Borrelia burgdorferi, the causative agent of Lyme disease. Previous studies from our group showed that BBK32 is a virulence factor in experimental Lyme disease and located the Fn-binding region to residues 21–205 of the lipoprotein.Methodology/Principal Findings
Studies aimed at identifying interacting sites between BBK32 and Fn revealed an interaction between the MSCRAMM and the Fn F3 modules. Further analysis of this interaction showed that BBK32 can cause the aggregation of human plasma Fn in a similar concentration-dependent manner to that of anastellin, the superfibronectin (sFn) inducing agent. The resulting Fn aggregates are conformationally distinct from plasma Fn as indicated by a change in available thermolysin cleavage sites. Recombinant BBK32 and anastellin affect the structure of Fn matrices formed by cultured fibroblasts and inhibit endothelial cell proliferation similarly. Within BBK32, we have located the sFn-forming activity to a region between residues 160 and 175 which contains two sequence motifs that are also found in anastellin. Synthetic peptides mimicking these motifs induce Fn aggregation, whereas a peptide with a scrambled sequence motif was inactive, suggesting that these motifs represent the sFn-inducing sequence.Conclusions/Significance
We conclude that BBK32 induces the formation of Fn aggregates that are indistinguishable from those formed by anastellin. The results of this study provide evidence for how bacteria can target host proteins to manipulate host cell activities. 相似文献74.
Patrick L Cable J Potts D Ignace D Barron-Gafford G Griffith A Alpert H Van Gestel N Robertson T Huxman TE Zak J Loik ME Tissue D 《Oecologia》2007,151(4):704-718
Global climate models predict that in the next century precipitation in desert regions of the USA will increase, which is
anticipated to affect biosphere/atmosphere exchanges of both CO2 and H2O. In a sotol grassland ecosystem in the Chihuahuan Desert at Big Bend National Park, we measured the response of leaf-level
fluxes of CO2 and H2O 1 day before and up to 7 days after three supplemental precipitation pulses in the summer (June, July, and August 2004).
In addition, the responses of leaf, soil, and ecosystem fluxes of CO2 and H2O to these precipitation pulses were also evaluated in September, 1 month after the final seasonal supplemental watering event.
We found that plant carbon fixation responded positively to supplemental precipitation throughout the summer. Both shrubs
and grasses in watered plots had increased rates of photosynthesis following pulses in June and July. In September, only grasses
in watered plots had higher rates of photosynthesis than plants in the control plots. Soil respiration decreased in supplementally
watered plots at the end of the summer. Due to these increased rates of photosynthesis in grasses and decreased rates of daytime
soil respiration, watered ecosystems were a sink for carbon in September, assimilating on average 31 mmol CO2 m−2 s−1 ground area day−1. As a result of a 25% increase in summer precipitation, watered plots fixed eightfold more CO2 during a 24-h period than control plots. In June and July, there were greater rates of transpiration for both grasses and
shrubs in the watered plots. In September, similar rates of transpiration and soil water evaporation led to no observed treatment
differences in ecosystem evapotranspiration, even though grasses transpired significantly more than shrubs. In summary, greater
amounts of summer precipitation may lead to short-term increased carbon uptake by this sotol grassland ecosystem. 相似文献
75.
Nesbitt TL Patel PA Yost MJ Goodwin RL Potts JD 《In vitro cellular & developmental biology. Animal》2007,43(1):10-16
Coronary vascular disease is one of the leading causes of mortality and morbidity in the United States. Therefore, a mechanistic
understanding of coronary vessel morphogenesis would aid in the innovation of new therapies targeting vascular disorders.
Moreover, a functionally equivalent in vitro model system allows for the delineation of the molecular mechanisms that regulate
coronary vessel development. In this study, we present a novel in vitro model system. This three-dimensional (3-D) model system
consists of a tubular scaffold, which is engineered from type-I collagen and has been optimized to support the growth of embryonic
cardiac tissues. In this report, proepicardial (PE) cells, the developmental precursors of coronary vessels, have been isolated
from several model species and cultured on this scaffold. In this model system, the PE cells were able to recapitulate several
aspects of coronary vessel morphogenesis including epicardial formation, the epicardial to mesenchymal transformation, and
de novo coronary vessel development or vasculogenesis. The differentiation of PE cells was characterized using a variety of
specific protein markers. The potential uses of this novel coronary developmental model are discussed. 相似文献
76.
Major histocompatibility complex heterozygosity reduces fitness in experimentally infected mice 总被引:2,自引:0,他引:2
It is often suggested that heterozygosity at major histocompatibility complex (MHC) loci confers enhanced resistance to infectious diseases (heterozygote advantage, HA, hypothesis), and overdominant selection should contribute to the evolution of these highly polymorphic genes. The evidence for the HA hypothesis is mixed and mainly from laboratory studies on inbred congenic mice, leaving the importance of MHC heterozygosity for natural populations unclear. We tested the HA hypothesis by infecting mice, produced by crossbreeding congenic C57BL/10 with wild ones, with different strains of Salmonella, both in laboratory and in large population enclosures. In the laboratory, we found that MHC influenced resistance, despite interacting wild-derived background loci. Surprisingly, resistance was mostly recessive rather than dominant, unlike in most inbred mouse strains, and it was never overdominant. In the enclosures, heterozygotes did not show better resistance, survival, or reproductive success compared to homozygotes. On the contrary, infected heterozygous females produced significantly fewer pups than homozygotes. Our results show that MHC effects are not masked on an outbred genetic background, and that MHC heterozygosity provides no immunological benefits when resistance is recessive, and can actually reduce fitness. These findings challenge the HA hypothesis and emphasize the need for studies on wild, genetically diverse species. 相似文献
77.
Feeding sodium butyrate (0.25–1 mg/ml) to cultures of Salinispora tropica NPS21184 enhanced the production of salinosporamide B (NPI-0047) by 319% while inhibiting the production of salinosporamide
A (NPI-0052) by 26%. Liquid chromatography mass spectrometry analysis of the crude extract from the strain NPS21184 fed with
0.5 mg/ml sodium [U-13C4]butyrate indicated that butyrate was incorporated as a contiguous four-carbon unit into NPI-0047 but not into NPI-0052. Nuclear
magnetic resonance analysis of NPI-0047 and NPI-0052 purified from the sodium [U-13C4]butyrate-supplemented culture extract confirmed this incorporation pattern. The above finding is the first direct evidence
to demonstrate that the biosynthesis of NPI-0047 is different from NPI-0052, and NPI-0047 is not a precursor of NPI-0052. 相似文献
78.
Highly frugivorous primates like chimpanzees (Pan trogolodytes) must contend with temporal variation in food abundance and quality by tracking fruit crops and relying more on alternative foods, some of them fallbacks, when fruit is scarce. We used behavioral data from 122 months between 1995 and 2009 plus 12 years of phenology records to investigate temporal dietary variation and use of fallback foods by chimpanzees at Ngogo, Kibale National Park, Uganda. Fruit, including figs, comprised most of the diet. Fruit and fig availability varied seasonally, but the exact timing of fruit production and the amount of fruit produced varied extensively from year to year, both overall and within and among species. Feeding time devoted to all major fruit and fig species was positively associated with availability, reinforcing the argument that chimpanzees are ripe fruit specialists. Feeding time devoted to figs-particularly Ficus mucuso (the top food)--varied inversely with the abundance of nonfig fruits and with foraging effort devoted to such fruit. However, figs contributed much of the diet for most of the year and are best seen as staples available most of the time and eaten in proportion to availability. Leaves also contributed much of the diet and served as fallbacks when nonfig fruits were scarce. In contrast to the nearby Kanywara study site in Kibale, pith and stems contributed little of the diet and were not fallbacks. Fruit seasons (periods of at least 2 months when nonfig fruits account for at least 40% of feeding time; Gilby & Wrangham., Behavioral Ecology and Sociobiology 61:1771-1779, 2007) were more common at Ngogo than Kanyawara, consistent with an earlier report that fruit availability varies less at Ngogo [Chapman et al., African Journal of Ecology 35:287-302, 1997]. F. mucuso is absent at Kanyawara; its high density at Ngogo, combined with lower variation in fruit availability, probably helps to explain why chimpanzee population density is much higher at Ngogo. 相似文献
79.
Ruth Joy Mitchell Adian M. Keith Jackie M. Potts Jasmine Ross Eileen Reid Lorna A. Dawson 《Plant and Soil》2012,352(1-2):65-84
Aim
To test if there is an interactive effect between tree and understory species on the soil microbial community (SMC), community level physiological profiles (CLPP) and soil micro-fauna.Method
A replicate pot experiment with five sapling tree species (Betula pendula, Betula pubescens, Sorbus aucuparia, Quercus petraea and Pinus sylvestris) and a no-tree treatment with and without Calluna vulgaris was established. After 21?months samples were taken for phospholipid fatty acid (PLFA) analysis, CLPP and soil microfauna assessment.Results
There was an interactive effect of tree species and Calluna on the SMC, CLPP and nematode densities. Calluna addition changed the SMC composition (increase in fungal PLFAs) and the CLPP (lower utilisation of most carbon sources but greater utilisation of phenolic acids). A multivariate test for homogeneity of dispersion showed that while Calluna addition resulted in the presence of an altered microbial composition, it did not result in there being less variability among the samples with Calluna than among the samples without Calluna. Sapling trees with Calluna present grew less well than trees without Calluna. Structural equation modelling showed that it is possible that Calluna had an indirect effect on the SMC via below-ground tree biomass as well as a direct effect.Conclusion
Interactions between trees and understory vegetation can impact on the composition of soil biota and their activity. 相似文献80.
Gunnarsson TG Sutherland WJ Alves JA Potts PM Gill JA 《Proceedings. Biological sciences / The Royal Society》2012,279(1727):411-416
The capacity of species to track changing environmental conditions is a key component of population and range changes in response to environmental change. High levels of local adaptation may constrain expansion into new locations, while the relative fitness of dispersing individuals will influence subsequent population growth. However, opportunities to explore such processes are rare, particularly at scales relevant to species-based conservation strategies. Icelandic black-tailed godwits, Limosa limosa islandica, have expanded their range throughout Iceland over the last century. We show that current male morphology varies strongly in relation to the timing of colonization across Iceland, with small males being absent from recently occupied areas. Smaller males are also proportionately more abundant on habitats and sites with higher breeding success and relative abundance of females. This population-wide spatial structuring of male morphology is most likely to result from female preferences for small males and better-quality habitats increasing both small-male fitness and the dispersal probability of larger males into poorer-quality habitats. Such eco-evolutionary feedbacks may be a key driver of rates of population growth and range expansion and contraction. 相似文献